Assessing Class 1 Integron Presence in Poultry Litter Amended with Wood Biochar and Wood Vinegar.

Class 1 integrons are mobile genetic elements that facilitate the spread of antibiotic resistance genes among bacteria. The use of prophylactic antibiotics has resulted in the rise of antibiotic resistance genes accumulating in a wide range of settings, including poultry houses and the agricultural fields where poultry litter is applied as a fertilizer. Biochar and wood vinegar are forest products wastes that have generated increasing attention as additives to agricultural soils. The objectives of this study were to observe the prevalence of class 1 integrons in poultry litter blended with biochar and wood vinegar over time and to verify a modified class 1 integron screening assay. Poultry litter blends were sampled and screened for class 1 integrons using polymerase chain reaction, and 80 products, 79 of which showed positive, were sent for DNA sequencing. The GenBank® BLAST database was used to verify the presence of the class 1 integron-integrase gene (intI1). There was no change in prevalence over time in poultry litter blends. Out of 79 PCR products that were intI1 positive, 78 showed at least 95% sequence identity to intI1 encoding bacteria and 64 showed at least 97% sequence identity. This indicates that this method was effective for conducting baseline surveillance of class 1 integrons in poultry litter and poultry litter-blended biochar and/or wood vinegar. Most significantly, class 1 integron prevalence did not decrease over time, further supporting the recalcitrance of these elements and the need for improved monitoring systems.

Effects of Loblolly Pine Biochar and Wood Vinegar on Poultry Litter Nutrients and Microbial Abundance.

Biochar, wood vinegar, and poultry litter are waste streams that can be utilized as soil amendments and fertilizers. However, poultry litter releases several pollutants through nutrient leaching and carries heavy microbial loads, including potential human pathogens. Improving nutrient retention and reducing microbial load in poultry litter may help protect environmental and human health and improve its value as a soil amendment. The objectives of this study were to determine how blending varying proportions of loblolly pine (Pinus taeda L.) biochar, wood vinegar, and poultry litter affected nutrient profiles and microbial abundance over time. Biochar inclusion rates were 0%, 5%, 10%, and 20%, and wood vinegar was applied at 2% w/w. Samples were taken at Day 0, 57, and 112 to measure nitrogen, phosphorus, potassium, pH, total fungi, and total bacteria. Nutrient levels generally decreased with increasing biochar level; however, biochar inclusion rates of 10% and 20% retained nitrogen and phosphorus and exhibited improved physical properties. Overall, adding wood vinegar decreased nutrient concentrations and showed a biocidal effect for bacteria and fungi. Bacteria and fungi showed different relationships with biochar inclusion rates, with fungi preferring higher biochar inclusion rates and bacteria flourishing at lower biochar inclusion rates.

Fungal Communities and Functional Guilds Shift Along an Elevational Gradient in the Southern Appalachian Mountains.

Nitrogen deposition alters forest ecosystems particularly in high elevation, montane habitats where nitrogen deposition is greatest and continues to increase. We collected soils across an elevational (788-1940 m) gradient, encompassing both abiotic (soil chemistry) and biotic (vegetation community) gradients, at eight locations in the southern Appalachian Mountains of southwestern North Carolina and eastern Tennessee. We measured soil chemistry (total N, C, extractable PO4, soil pH, cation exchange capacity [ECEC], percent base saturation [% BS]) and dissected soil fungal communities using ITS2 metabarcode Illumina MiSeq sequencing. Total soil N, C, PO4, % BS, and pH increased with elevation and plateaued at approximately 1400 m, whereas ECEC linearly increased and C/N decreased with elevation. Fungal communities differed among locations and were correlated with all chemical variables, except PO4, whereas OTU richness increased with total N. Several ecological guilds (i.e., ectomycorrhizae, saprotrophs, plant pathogens) differed in abundance among locations; specifically, saprotroph abundance, primarily attributable to genus Mortierella, was positively correlated with elevation. Ectomycorrhizae declined with total N and soil pH and increased with total C and PO4 where plant pathogens increased with total N and decreased with total C. Our results demonstrate significant turnover in taxonomic and functional fungal groups across elevational gradients which facilitate future predictions on forest ecosystem change in the southern Appalachians as nitrogen deposition rates increase and regional temperature and precipitation regimes shift.