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IntroductionPromoting sustainable crop production is enhanced by an effective method to assess soil health. However, soil health assessment is challenging due to multiple interactions among dynamic soil properties (i.e., soil health indicators) across management practices and agroecological regions. We tested several currently popular soil health assessment methods for cropping systems in Tennessee in the southeastern US and found that these methods failed to differentiate Tennessee soil health under long-term conservation and conventional management.Materials and methodsThis study developed a Tennessee weighted soil health index (WSHI) by: 1) selecting a set of management-sensitive soil health indicators, 2) assigning meaningful weights to indicators, and 3) normalizing the scores based on regionally relevant undisturbed natural reference sites. The tested cropping systems treatments were moldboard plow (MP) in continuous soybean (SS), no tillage (NT) in SS, NT with wheat cover (NTW) in SS, no cover and chisel plow (NCCT) in continuous cotton (CC), no cover and no tillage (NCNT) in CC, and hairy vetch cover and no tillage (VCNT) in CC. In addition, two woodlots and one grassland sites in the vicinity of the cropping systems were selected to represent undisturbed natural systems.Results and discussionOut of 22 indicators that proved to be management-sensitive, six were selected as a minimum dataset (MDS). These were particulate organic matter C (POM-C), soil respiration from 4-day incubation (4d CO2), small macroaggregate (0.250-2mm)-associated C (SMA-C), surface hardness (PR15), microbial biomass N (MBN), and bulk density (BD). Measured values of the MDS indicators were transformed into unitless normalized scores (based on the regional range of the indicator), and finally integrated into WSHI scores using a weighted-addition approach. Additionally, the soil health gap (SHG) between the soil health of the regional reference system and different cropping systems was calculated. Results revealed that WSHI strongly differentiated soil health between long-term conservation and conventional managements practices. The WSHI scores for southeastern cropland soils varied as follows: VCNT = NTW > NT > NCNT ≥ NCCT ≥ MP. The SHGs under MP, NCCT, NCNT, NT, NTW, and VCNT were 85.5, 79.9, 68, 45.1, 25.2, and 24.3, respectively, relative to the average WSHI of three undisturbed systems. Results showed that the WSHI approach is effective in more meaningful regional assessment of soil health and SHG can be a potential metric for comparing soil health across agroecological regions.

Home gardeners’ concerns for the environment are expressed both in the ecofriendly gardening practices they use and in environmental attributes they prefer in the gardening products they purchase. This study uses data from a 2018 survey of 601 Tennessee outdoor home gardeners and a multiple indicators multiple causes (MIMIC) model to illustrate how outdoor home gardener demographics, expenditures, information use, and attitudes influence use of ecofriendly gardening practices and preferences for environmental attributes in home gardening supplies. Practices considered include planting pollinator plants, using rainwater collectors, composting, recycling gardening supplies packaging, using organic gardening methods, and use of soil testing. Gardening supply product attributes include decreased need for fertilizer, pesticides, and water; native plant species; organically produced products; and recyclable packaging. The most widely used practice is recycling gardening supplies packaging, and the least used is soil testing. Gardeners with a greater propensity to use the six gardening practices include male, college graduates, who spend relatively more of their income on gardening supplies, and consider themselves as being knowledgeable about environmental issues. The gardening supply product attribute most widely considered as important is decreased need for pesticides, and least widely considered as important are native species and organically produced. Gardeners more likely to prefer the six gardening supply product attributes include older gardeners, who seek other gardeners for information, and who perceived themselves as being knowledgeable about the environment. This same group likes to grow their own food and feels responsibility for protecting the environment for future generations.

BackgroundTreatment burden is the effort required of patients to look after their health and the impact this has on their functioning and wellbeing. It is likely treatment burden changes over time as circumstances change for patients and health services. However, there are a lack of population-level studies of treatment burden change and factors associated with this change over time. Furthermore, there are currently no practical screening tools for treatment burden in time-pressured clinical settings or at population level.Methods and analysisThis is a three-year follow-up of a cross-sectional survey of 723 people with multimorbidity (defined as three or more long-term conditions; LTCs) registered at GP practices in in Dorset, England. The survey will repeat collection of information on treatment burden (using the 10-item Multimorbidity Treatment Burden Questionnaire (MTBQ) and a novel single-item screening tool), sociodemographics, medications, LTCs, health literacy and financial resource, as at baseline. Descriptive statistics will be used to compare change in treatment burden since the baseline survey in 2019 and associations of treatment burden change will be assessed using regression methods. Diagnostic test accuracy metrics will be used to evaluate the single-item treatment burden screening tool using the MTBQ as the gold-standard. Routine primary care data (including demographics, medications, LTCs, and healthcare usage data) will be extracted from medical records for consenting participants. A forward-stepwise, likelihood-ratio logistic regression model building approach will be employed in order to assess the utility of routine data metrics in quantifying treatment burden in comparison to self-reported treatment burden using the MTBQ.ImpactTo the authors' knowledge, this will be the first study investigating longitudinal aspects of treatment burden. Findings will improve understanding of the extent to which treatment burden changes over time for people with multimorbidity and factors contributing to this change, as well as allowing better identification of people at risk of high treatment burden.

Abstract Seasonal flooding following heavy rain deposits large amounts of sediments on productive lands in the lower parts of fields in western Tennessee. The deposited sediments have high proportion of sand particles that negatively affect soil physiochemical properties, which make the soil uneconomic for farming. Soil amendments such as biochar, a by‐product of renewable energy production from organic waste materials, have the potential to remediate sandy soil left after flood events and improve crop yields by increasing water holding capacity and soil nutrient content. The objective of this project was to evaluate the effect of biochar produced from two types of hardwood feedstocks on water retention and corn (Zea mays L.) growth in sandy soil. A greenhouse experiment quantifying the growth of corn was conducted in a randomized complete block design with five replications. Sandy soil was amended with three biochar rates (0, 10, and 20% by volume) under two irrigation levels. The irrigation levels were control and a dry treatment based on the past 10 yr of rainfall data. Biochar application greatly improved water retention in the flooded sandy soil. Biochar increased soil K and P concentration. However, at the end of the study, corn growth was not different in biochar amended and non‐amended sandy soil. This research demonstrated that biochar as a soil amendment has the potential to improve quality attributes of poor soil, such as the soil water and nutrient concentration in a previously flooded sandy soils.

Previous land surface modeling efforts to predict and understand water budgets in the U.S. Southeast for soil water management have struggled to characterize parts of the region due to an extensive presence of fragipan soils for which current calibration approaches are not adept at handling. This study presents a physically based approach for calibrating fragipan-dominated regions based on the “effective” soil moisture capacity concept, which accounts for the dynamic perched saturation zone effects created by the low hydraulic capacities of the fragipan layers. The approach is applied to the Variable Infiltration Capacity model to develop a hydrologic model of the Obion River Watershed (ORW), TN, which has extensive fragipan coverage. Model calibration was performed using observed streamflow data, as well as evapotranspiration and soil moisture data, to ensure correct partitioning of surface and subsurface fluxes. Estimated Nash-Sutcliffe coefficients for the various sub-drainage areas within ORW were all greater than 0.65, indicating good model performance. The model results suggest that ORW has a high responsivity and high resilience. Despite forecasted temperature increases, the simulation results suggest that water budget trends in the ORW are unlikely to change significantly in the near future up to 2050 due to sufficient precipitation amounts.

Spiroplasma, small motile wall-less bacteria, are linked by molecular and serological studies to the transmissible spongiform encephalopathies (TSEs), which include scrapie in sheep, chronic wasting disease (CWD) in deer and Creutzfeldt-Jakob disease in humans. In this study, two experiments were undertaken to determine the role of spiroplasma in the pathogenesis of TSE. In experiment 1, Spiroplasma mirum, a rabbit tick isolate that had previously been shown to experimentally induce spongiform encephalopathy in rodents, was inoculated intracranially (IC) into ruminants. S. mirum-inoculated deer manifested clinical signs of TSE after 1.5 to 5.5 months incubation. The deer, as well as sheep and goats, inoculated with S. mirum developed spongiform encephalopathy in a dose-dependent manner. In experiment 2, spiroplasma closely related to S. mirum were isolated from TSE-affected brains via passage in embryonated eggs, and propagated in cell-free M1D media. Spiroplasma spp. isolates from scrapie-affected sheep brain and from CWD-affected deer brain inoculated IC into sheep and goats induced spongiform encephalopathy closely resembling natural TSE in these animals. These data show spiroplasma to be consistently associated with TSE, and able experimentally to cause TSE in ruminant animal models, therein questioning the validity of studies that have concluded the prion, a miss-folded protease-resistant protein that builds up in TSE brains during the course of the disease, to be the sole causal agent. The spiroplasma infection models reported here will be important for investigating factors involved in the pathogenesis of TSE since ruminants are the natural hosts.

Swine erysipelas vaccines are routinely used to protect pigs against peracute and acute/urticarial forms of Erysipelothrix. Between 1995 and 1998, 34 swine herds across four Australian states experienced vaccine failure. Forty-four isolates of Erysipelothrix rhusiopathiae of serovars 2, 1a, 1b and 1bx21 were recovered from 15 of these 34 vaccine breakdown herds. These isolates were characterised by restriction fragment length polymorphism (RFLP) analyses using RsaI and AluI on whole cell DNA and for the presence of plasmid DNA. Results were compared with those of 20 isolates from 16 herds unaffected by vaccine breakdown and 13 isolates representing 10 reference strains. The majority of breakdown herds possessed isolates of serovar 2 (9/15 herds), followed by serovar 1a (5 herds). No geographic predominance of a single serovar was evident. The identification of 10 RsaI profiles from whole cell DNA among the 44 isolates from 15 breakdown herds indicated that a single, new clonal lineage of E. rhusiopathiae was not responsible for vaccine failure. RsaI RFLP analyses detected a further 14 distinct profiles among 20 field strains unassociated with vaccine breakdowns, and none matched profiles of the 10 serovar reference strains for serovars 1a, 1b, 2 or 21. This technique is recommended for epidemiological studies of E. rhusiopathiae strains.

A comprehensive genetic analysis of 60 Mycoplasma sp. bovine group 7 isolates from different geographic origins and epidemiological settings is presented. Twenty-four isolates were recovered from the joints of calves during sporadic episodes of polyarthritis in geographically distinct regions of Queensland and New South Wales, Australia, including two clones of the type strain PG5O. A further three Australian isolates were also recovered from the tympanic bulla, retropharyngeal lymph node and the lung and another three isolates had unconfirmed histories. Six isolates originated from Germany, Portugal, Nigeria, and France. Twenty-four epidemiologically related isolates of Mycoplasma sp. bovine group 7 were recovered from multiple tissue sites and body fluids of infected calves with polyarthritis, mastitic milk, and from the stomach contents, lung and liver from aborted foetuses in three large, centrally managed dairy herds in New South Wales, Australia. Restriction endonuclease analysis (REA) of genomic DNA differentiated 29 Cfol profiles among these 60 isolates and grouped all 24 epidemiologically related isolates in a defined pattern showing a clonal origin. Three isolates of this clonal cluster were recovered from mastitic milk and the synovial exudate of clinically-affected calves and appeared sporadically for periods up to 18 months after the initial outbreak of polyarthritis indicating a persistent, close association of the organism with cattle in these herds. The Cfol profile representative of the clonal cluster was distinguishable from profiles of isolates recovered from multiple, unrelated cases of polyarthritis in Queensland and New South Wales and from other countries. All 24 isolates from the clonal cluster possessed a plasmid (pBG7AU) with a molecular size of 1022 bp. DNA sequence analysis of pBG7AU identified two open reading frames sharing 81 and 99% DNA sequence similarity with hypothetical replication control proteins A and B respectively, previously described in plasmid pADB201 isolated from M. mycoides subspecies mycoides. Other isolates of bovine group 7, epidemiologically unrelated to the clonal cluster, including two clones of the type strain PG5O, possessed a similar-sized plasmid. These data confirm that Mycoplasma sp. bovine group 7 is capable of migrating to, and multiplying within, different tissue sites within a single animal and among different animals within a herd.

The apxIVA gene, a recently discovered RTX determinant of Actinobacillus pleuropneumoniae, was shown to be species-specific. DNA hybridization experiments using probes for various regions of apxIVA revealed that the 3'-terminus of this gene was present in all 14 serotypes of A. pleuropneumoniae but absent from phylogenetically related species. A primer pair spanning this region specifically amplified a 422bp fragment in PCR experiments with DNA from the reference strains of the 14 serotypes and 194 field strains isolated from various geographic locations worldwide. DNA sequence analysis of PCR products derived from all serotypes were identical except in serotypes 3, 8, and 10, which showed minor differences. The PCR did not amplify any product when DNA from 17 different bacterial species closely related to A. pleuropneumoniae was used as template. In addition, the PCR was negative with DNA of several Actinobacillus sp. which were initially characterized as A. pleuropneumoniae using routine phenotypic and serological analyses but which were subsequently shown by 16S rRNA sequence analysis to belong to yet undefined Actinobacillus species. The sensitivity of the PCR was determined to be 10pg of A. pleuropneumoniae DNA. A set of nested primers amplified a 377bp fragment specifically with A. pleuropneumoniae DNA. DNA titration experiments using the flanking and nested primer pairs showed an improved level of sensitivity to approximately 10fg of genomic DNA. The nested PCR was used to monitor the spread of A. pleuropneumoniae in pigs experimentally infected with a virulent serotype 1 strain and housed in a controlled environment facility. A. pleuropneumoniae DNA could be detected by nested PCR in nasal swab samples of infected pigs receiving either a high dose (5x10(5)) or a low dose (1x10(4)) challenge and in unchallenged cohorts that were contact-infected by the inoculated animals. Furthermore, PCR confirmed the presence of A. pleuropneumoniae in 16/17 homogenates from necrotic lung lesions, while the bacterium was successfully recovered from 13 of these lesions by culture.

Twenty-five unique CfoI-generated whole-cell DNA profiles were identified in a study of 30 Paenibacillus alvei isolates cultured from honey and diseased larvae collected from honeybee (Apis mellifera) colonies in geographically diverse areas in Australia. The fingerprint patterns were highly variable and readily discernible from one another, which highlighted the potential of this method for tracing the movement of isolates in epidemiological studies. 16S rRNA gene fragments (length, 1,416 bp) for all 30 isolates were enzymatically amplified by PCR and subjected to restriction analysis with DraI, HinfI, CfoI, AluI, FokI, and RsaI. With each enzyme the restriction profiles of the 16S rRNA genes from all 30 isolates were identical (one restriction fragment length polymorphism [RFLP] was observed in the HinfI profile of the 16S rRNA gene from isolate 17), which confirmed that the isolates belonged to the same species. The restriction profiles generated by using DraI, FokI, and HinfI differentiated P. alvei from the phylogenetically closely related species Paenibacillus macerans and Paenibacillus macquariensis. Alveolysin gene fragments (length, 1, 555 bp) were enzymatically amplified from some of the P. alvei isolates (19 of 30 isolates), and RFLP were detected by using the enzymes CfoI, Sau3AI, and RsaI. Extrachromosomal DNA ranging in size from 1 to 10 kb was detected in 17 of 30 (57%) P. alvei whole-cell DNA profiles. Extensive biochemical heterogeneity was observed among the 28 P. alvei isolates examined with the API 50CHB system. All of these isolates were catalase, oxidase, and Voges-Proskauer positive and nitrate negative, and all produced acid when glycerol, esculin, and maltose were added. The isolates produced variable results for 16 of the 49 biochemical tests; negative reactions were recorded in the remaining 30 assays. The genetic and biochemical heterogeneity in P. alvei isolates may be a reflection of adaptation to the special habitats in which they originated.

Melissococcus pluton, the causative agent of European foulbrood is an economically significant disease of honey bees (Apis mellifera) across most regions of the world and is prevalent throughout most states of Australia. 49 Isolates of M. pluton recovered from diseased colonies or honey samples in New South Wales, Queensland, South Australia, Tasmania and Victoria were compared using SDS-PAGE, Western immunoblotting and restriction endonuclease analyses. DNA profiles of all 49 geographically diverse isolates showed remarkably similar AluI profiles although four isolates (one each from Queensland, South Australia, New South Wales and Victoria) displayed minor profile variations compared to AluI patterns of all other isolates. DNA from a subset of the 49 Australian and three isolates from the United Kingdom were digested separately with the restriction endonucleases CfoI, RsaI and DraI. Restriction endonuclease fragment patterns generated using these enzymes were also similar although minor variations were noted. SDS-PAGE of whole cell proteins from 13 of the 49 isolates from different states of Australia, including the four isolates which displayed minor profile variations (AluI) produced indistinguishable patterns. Major immunoreactive proteins of approximate molecular masses of 21, 24, 28, 30, 36, 40, 44, 56, 60, 71, 79 and 95 kDa were observed in immunoblots of whole cell lysates of 22 of the 49 isolates and reacted with rabbit hyperimmune antibodies raised against M. pluton whole cells. Neither SDS-PAGE or immunoblotting was capable of distinguishing differences between geographically diverse isolates of M. pluton. Collectively these data confirm that Australian isolates of M. pluton are genetically homogeneous and that this species may be clonal. Plasmid DNA was not detected in whole cell DNA profiles of any isolate resolved using agarose gel electrophoresis.