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The aim of this study was to evaluate the treatment effects of Ca(OH)2 solutions applied at different concentrations (2.5%, 5%, and 7.5%) on the structural and chemical composition of sugarcane bag...

Saprolite, weathered bedrock, is being used to dispose of domestic sewage through septic system drainfields, but its ability to remove coliforms is unknown. This study determined if Escherichia coli could be removed by a sandy loam saprolite material. Triplicate columns containing saprolite were prepared with lengths of 30, 45, and 60 cm. A 215-mL solution containing 1 × 105 CFU/100 mL of non-toxic E. coli was applied to the top of each column for 5 days/week for 13 weeks, and selected outflow samples were analyzed for E. coli. Control columns had only tap water applied to them at the same time. Significantly higher (p ≤ 0.10 compared to controls) E. coli concentrations were only detected in samples collected at the end of week 3 for the 30-cm columns and week 4 for the 45-cm columns. E. coli concentrations were small and ranged from approximately 2 to 3 MPN/100 mL. No E. coli were detected in any outflow from the 60-cm columns. From weeks 5 to 13, E. coli concentrations from all columns were either undetectable or not significantly different from the control. The results showed that 60 cm of sandy loam saprolite was sufficient for the removal of E. coli from simulated wastewater.

Interactions between above- and below-ground monoculture forest plantation components are critical to tree growth and development. Within the Central Hardwoods Region (CHR), synergistic relationships between tree species and soil microbial community structure and function have received limited research attention. Soil microbes are integral to forest ecosystems as their activities intrinsically promote soil organic matter decomposition, nutrient cycling, and ecosystem functioning. Here, we examined soils from two perfectly aligned stands of black walnut (BW, Juglans nigra L.) and Northern red oak (RO, Quercus rubra L.) trees. Measurements of selected soil chemical properties, microbial community structure using ester-linked fatty acid methyl ester (EL-FAME), and soil enzyme activities (EAs) were used. Analysis of modifications within microbial communities showed a significant positive response to BW based upon soil EAs and microbial indicators, compared to RO. Seasonal comparisons predictably revealed higher microbial activities during summer. Fungi dominated the soil microbial community structure with a fungal/bacterial ratio of 2:1. Gram-positive rather than Gram-negative bacteria or actinomycetes dominated the bacterial community. The activity of the soil enzymes ß-glucosidase and arylsulfatase increased, but ß-glucosaminidase and acid phosphatase decreased. Additionally, acid phosphatase and arbuscular mycorrhizal fungi revealed strong correlations. The differences observed in biological properties, specifically microbial communities and EAs, highlight the varied responses to BW and RO soil biology and subsequent soil ecosystem functions. These results indicate that variations in microbial abundance and soil functions occur throughout the course of an entire year.

International audience; A good bioindicator of soil health is one that perceives minor changes due to changes in soil management and leads to changes in the processes of the soil ecosystem. The aim was to evaluate the use of soil bioindicators in the determination of the utility of two different Cedrela odorata forestry management systems. The litter and soil samples were collected from the rhizosphere of a monoculture (with no fertilization nor irrigation), and a co-culture (with fertilization and irrigation) stand with Citrus latifolia. The soil and litter bioindicators measured were the activity of acetylesterase (FDA), acid phosphatase (AcPh), alkaline phosphatase (AlkPh), laccase, and the estimation of the potential net rate of carbon mineralization (PNRCM). The results indicated that soil enzyme activities (FDA and phosphatases) in the litter were sensitive to the different management system treatments. Enzymatic activity, per unit dry weight, was higher in litter of co-culture (FDA = 1.05 nkat g-1 AcPh = 1.33 nkat g-1; AlkPh = 03.0 nkat g-1) than in litter of monoculture (FDA = 0.617nkat g-1; AcPh = 0.40 nkat g-1; AlkPh = 0. 983 nkat g-1). Differences in soil total nitrogen content, organic matter (OM) content, and micronutrients were found. In general, the co-culture stand (with fertilization and irrigation) with C. latifolia showed the best results.

Biochar offers several benefits as a soil amendment, including increased soil fertility, carbon sequestration, and water-holding capacity in nutrient-poor soils. In this study, soil samples with and without biochar additives were collected for two consecutive years from an experimental field plot to examine its effect on the microbial community structure and functions in sandy soils under peach-trees (Prunus persica). The four treatments evaluated consisted of two different rates of biochar incorporated into the soil (5%, and 10%, v/v), one “dynamic” surface application of biochar, and a 0% biochar control. Fatty acid methyl ester (FAME) analysis was used to assess the microbial community structure, and enzyme activities involved in C, N, P, and S nutrient cycling were used as a means of assessing soil functionality. Total FAME and bacterial indicators increased by 18% and 12%, respectively, in the 10% incorporated and 5% surface applied treatments. Biochar applications increased β-glucosaminidase and arylsulfatase activities, 5–30% and 12–46%, respectively. β-glucosidase and acid phosphatase activities decreased by approximately 18–35% and 5–22% in the 0–15 cm soils. The overall results suggest that biochar’s addition to the sandy soils stimulated microbial activity, contributing to the increased mean weight diameter (MWD), C sequestration, and consequential soil health. The changes in microbial community structure and functions may be useful predictors of modifications in soil organic matter (SOM) dynamics due to the long-term application of pine biochar in these systems.

Sulfatases which cleave sulfate esters in biological systems are key enzymes that deserve special attention due to their significant roles in organic sulfur (OS) mineralization and inorganic sulfur () release. In this study, in-vitro experiments were conducted to evaluate S bonded substrate hydrolysis by a commercially available arylsulfatase (EC 3.1.6.1) from Aerobacter aerogenes. The enzyme-substrate interactions were assessed to determine: 1) rate of hydrolysis, 2) catalytic efficiency, 3) thermal stability, and 4) optimal pH of this enzyme. Arylsulfatase exhibited substrate hydrolysis with a high affinity for p-nitrophenyl sulfate (potassium 4-nitrophenyl sulfate (pNPS)). The optimum activity for the enzyme was observed to occur at a pH of 7.1. The optimal temperature was 37°C but ranged from 35°C - 45°C. The apparent Km and Kcat of the enzyme for pNPS hydrolysis at the optimal pH, and temperature were determined to be 1.03 mM and 75.73 μM/min, respectively. This work defines the catalytic and kinetic properties of arylsulfatase (EC 3.1.6.1) and confirms the optimal conditions for sulfatase activity testing. The resulting information is useful in elucidating the contributions that individual enzymes have for specific reactions rather than relying on traditional total enzyme activity measurements.

Forest floor and mineral soil manipulations influence the soil biogeochemical properties important for loblolly pine (Pinus taeda L.) tree growth. The impacts of forest floor manipulations on soil aggregate stability and the presence of ectomycorrhizal fungi (EMF), was assessed to elucidate the relationship between EMF abundance and aggregate stability. The study site consists of a 14-year-old loblolly pine plantation managed by Weyerhaeuser Company in the Lower Coastal Plain, approximately 8 miles east of New Bern, North Carolina, USA. The soil samples were collected from the top 7.62 cm of each soil treatment which includes three levels of forest floor retention: removed, control, and doubled and two levels of forest floor mixing with the mineral soil: mixed and unmixed. Ectomycorrhizal fungi abundance was evaluated by ester-linked fatty acid methyl ester analysis and microbial community functionality was assessed by acid-phosphatase activity measurement. Aggregate stability was assessed using the aggregate mean weight diameter approach. Results indicate that the forest floor manipulations had no significant impact on aggregate stability and EMF abundance. However, a positive relationship between EMF abundance and aggregate stability was identified. Removing the forest floor resulted in a soil bulk density increase of 0.18 g cm−3 compared to doubling the forest floor. Our results demonstrate that some mineral soil properties recover relatively quickly from forest floor manipulations. The study informs forest managers interested in how soil responds to forest floor manipulation and the interaction between EMF and aggregate stability.

Prescribed burning and tree thinning are commonly used restoration practices for US forests management to increase forest productivity and enhance plant and animal diversity. The impact of these practices in Alabama’s Bankhead National Forest (BNF) to soil microbial components and overall forest soil health are unknown. We hypothesized that microbial assemblages and enzyme activities are continuously changing in forest ecosystems especially due to management selections. Therefore, the objective of this study was to assess changes in microbial community compositions (fungal vs bacterial populations) via fatty acid methyl ester (FAME) profiling and several enzyme activities (β-glucosaminidase, acid phosphatase, arylsulfatase, β-glucosidase, xylanase, laccase, and manganese peroxidase) critical to soil organic matter (SOM) dynamics and biogeochemical cycling. In this forest, heavily-thinned plots without burning or less frequent burning treatments seemed to provide more favorable conditions (higher pH and lower C:N ratios) for C and N mineralization. This may explain a slight increase (by 12%) detected in fungi:bacteria (F:B) ratio in the heavily-thinned plots relative to the control. Thinned (lightly and heavily) plots showed greater ligninolytic (laccase and MnP) activities and lower β-glucosidase and β-glucosaminidase activities compared to the no-thinned plots probably due to increase depositions of woody recalcitrant C materials. We observed significant but negative correlations between the ligninolytic laccase and manganese peroxidase (Lac and MnP) enzymes respectively, with MBC (?0.45* and ?0.68** respectively) and MBN (?0.43* and ?0.65** respectively). Prescribed burning treatment reduced microbial biomass C and N of the 9-yr burned plot/lightly thinned plotsprobably due to depletion of labile C sources with the high temperatures, leaving mostly recalcitrant C sources as available soil substrates. Gram-positive bacteria (i15:0, a15:0, i17:0, and a17:0), actinomycetes (10-Me17:0, 10-Me18:0), AMF (16:1ω5c), and saprophytic fungi (18:1ω9c), largely contributed to the microbial compositions. This study bridges knowledge gaps in our understanding of microbial community compositions and enzyme-mediated processes in repeatedly burned and thinned forest ecosystems.

Manganese oxides, which may be biogenically produced in both pristine and contaminated environments, have a large affinity for many trace metals. In this study, water and Mn oxide-bearing biofilm samples were collected from the components of a pump and treat remediation system at a superfund site. To better understand the factors leading to their formation and their effects on potentially toxic metal fate, we conducted a chemical, microscopic, and spectroscopic characterization of these biofilm samples. Scanning electron microscopy revealed the presence of Mn oxides in close association with biological structures with morphologies consistent with fungi. X-ray absorption spectroscopy (XAS) and X-ray diffraction (XRD) revealed the oxides to be a mixture of layer and tunnel structure Mn(iv) oxides. In addition, XAS suggested that Ba, Co, and Zn all primarily bind to oxides in the biofilm in a manner that is analogous to synthetic or laboratory grown bacteriogenic Mn oxides. The results indicate that Mn oxides produced by organisms in the system may effectively scavenge metals, thus highlighting the potential utility of these organisms in designed remediation systems.

Saprolite, weathered bedrock, is being used to dispose of domestic sewage through septic system drainfields, but the thickness of saprolite needed to remove biological contaminants is unknown for most saprolites. This study developed and tested a simple method for estimating the thickness of saprolite needed below septic drainlines to filter E. coli from wastewater using estimates of the volume of pores that are smaller than the length of the coliform (≤10 μm). Particle size distribution (texture) and water retention data were obtained for 12 different saprolites from the Piedmont and Mountain regions of North Carolina (N.C.). Saprolite textures ranged from clay loam to coarse sand. The volume of pores with diameters ≤10 μm were determined by water retention measurements for each saprolite. The data were used in an equation to estimate the saprolite thickness needed to filter E. coli. The estimated saprolite thicknesses ranged from 36 cm in the clay loam to 113 cm for the coarse sand. The average thickness across all samples was 58 cm. Saprolite thickness estimates increased as silt percentage decreased and as sand percentage and in situ saturated hydraulic conductivity increased. Silt percentage may be most useful for estimating appropriate saprolite thicknesses in the field.

Manganese-oxidizing fungi support bioremediation through the conversion of manganese ions into manganese oxide deposits that in turn adsorb manganese and other heavy metal ions from the environment. Manganese-oxidizing fungi were immobilized onto nanofiber surfaces to assist remediation of heavy metal–contaminated water. Two fungal isolates, Coniothyrium sp. and Coprinellus sp., from a Superfund site (Lot 86, Farm Unit #1) water treatment system were incubated in the presence of nanofibers. Fungal hyphae had strong association with nanofiber surfaces. Upon fungal attachment to manganese chloride–seeded nanofibers, Coniothyrium sp. catalyzed the conformal deposition of manganese oxide along hyphae and nanofibers, but Coprinellus sp. catalyzed manganese oxide only along its hyphae. Fungi–nanofiber hybrids removed various heavy metals from the water. Heavy metal ions were adsorbed into manganese oxide crystalline structure, possibly by ion exchange with manganese within the manganese oxide. Hybrid materials of fungal hyphae and manganese oxides confined to nanofiber-adsorbed heavy metal ions from water.

Identification of microbial assemblages predominant under natural extreme climatic events will aid in our understanding of the resilience and resistance of microbial communities to climate change. From November 2010 to August 2011, the Southern High Plains (SHP) of Texas, USA, received only 39.6 mm of precipitation (vs. the historical average of 373 mm) and experienced the three hottest months (June–August 2011) since record keeping began in 1911. The objective of this study was to characterize soil bacterial (16 S rRNA gene) and fungal (internal transcribed spacer 1–4, ITS1-ITS4) species distribution and diversity via pyrosequencing during the peak of the drought/heat wave in July 2011 and when the Drought Index and temperatures were lower in March 2012. Samples were collected from two different soil types (loam and sandy loam) under two different dryland cropping histories (monoculture vs. rotation). Fungal Diversity Indexes were significantly higher after the drought/heat wave while Bacterial Indexes were similar. Bacterial phyla distribution in July 2011 was characterized by lower relative abundance of Acidobacteriaand Verrucomicrobia, and greater relative abundance of Proteobacteria, Chloroflexi, Actinobacteria and Nitrospirae than March 2012 samples. Further grouping of pyrosequencing data revealed approximately equal relative proportions of Gram positive (G+) and Gram negative (G−) bacteria in July 2011, while G− bacteria predominated in March 2012. Fungal class Dothideomycetes was approximately two times greater in July 2011 than in March 2012, while the class Sordariomycetes and a group of unidentified OTUs from Ascomycota increased from July 2011 to March 2012. Microbial community composition was less influenced by management history than by the difference in climatic conditions between the sampling times. Correspondence analysis identified assemblages of fungal and bacterial taxa associated with greater enzyme activities (EAs) of C, N, or P cycling found during the drought/heat wave. Microbial assemblages associated with arylsulfatase activity (key to S cycling), which increased after the drought/heat wave, were identified (Streptomyces parvisporogenes, Terrimonas ferruginea and Syntrophobacter sp.) regardless of the soil and management history. The distinct microbial composition found in July 2011 may represent assemblages essential to maintaining ecosystem function during extreme drought and intense heat waves in semiarid agroecosystems.

Soil organic matter (SOM) is essential for soil water holding capacity, aggregation, and biodiversity. Little information is available regarding the carbon (C) functional groups carried away in wind eroded sediments away from the source soil. Mid-infrared (MidIR) spectroscopy was used on wind tunnel-blown sediments eroded from a loam soil during the fallow period of different cropping systems and tillage managements in Akron, Colorado. The soil was managed as fallow-winter wheat ( Triticum aestivum L.) under conventional tillage (F–Wct) or no tillage (F–Wnt) and fallow–wheat–corn under no tillage (F–W–Cnt). Two wind eroded sediments were evaluated: fine dust ( −1 ), and clays (3690–3620 cm −1 ). The saltation-sized material showed higher absorbance for quartz from 2000–1800 cm −1 and reduced absorbance from 1250–1050 cm −1 . Both wind eroded sediments showed higher absorbance for –OH/NH groups and aliphatic CH from no-till soil. Finer dust sediments, which travel greater distances from the source soil than saltation size material, can carry away higher levels of aliphatic-carbon compounds and clays with potential negative impacts on SOM quantity and quality, and consequently the sustainability of these agroecosystems.

Extreme droughts and heat waves due to climate change may have permanent consequences on soil quality and functioning in agroecosystems. During November 2010 to August 2011, the Southern High Plains (SHP) region of Texas, U.S., a large cotton producing area, received only 39.6 mm of precipitation (vs. the historical avg. of 373 mm) and experienced the hottest summer since record keeping began in 1911. Several enzyme activities (EAs) important in biogeochemical cycling were evaluated in two soils (a loam and a sandy loam at 0–10 cm) with a management history of monoculture (continuous cotton) or rotation (cotton and sorghum or millet). Samplings occurred under the most extreme drought and heat conditions (July 2011), after precipitation resulted in a reduction in a drought severity index (March 2012), and 12 months after the initial sampling (July 2012; loam only). Eight out of ten EAs, were significantly higher in July 2011 compared to March 2012 for some combinations of soil type and management history. Among these eight EAs, enzymes key to C (β-glucosidase, β-glucosaminidase) and P cycling (phosphodiesterase, acid and alkaline phosphatases) were significantly higher (19–79%) in July 2011 than in March 2012 for both management histories regardless of the soil type ( P > 0.05). When comparing all sampling times, the activities of alkaline phosphatase, aspartase and urease (rotation only) showed this trend: July 2011 > March 2012 > July 2012. Activities of phosphodiesterase, acid phosphatase, α-galactosidase, β-glucosidase and β-glucosaminidase were higher in July 2011 than July 2012 in at least one of the two management histories. Total C was reduced significantly from July 2011 to March 2012 in the rotation for both soils. Only the activities of arylsulfatase (avg. 36%) and asparaginase showed an increase from July 2011 to March 2012 for both soil types, which may indicate they have a different origin/location than the other enzymes. EAs continued to be a fingerprint of the soil management history (i.e., higher EAs in the rotation than in monoculture) during the drought/heat wave. This study provided some of the first evidence of the adverse effects of a natural, extreme drought and heat wave on soil quality in agroecosystems as indicated by EAs involved in biogeochemical cycling.

A challenging aspect of educating secondary students is integrating complex scientific concepts related to modern research topics into lesson plans that students can relate to and understand at a basic level. One method of encouraging the achievement of learning outcomes is to use real-world applications and current research to fuel student interest and excitement. This approach can be reinforced by use of demonstrations, which spur in-class discussions and can be integrated into out-of-class assignments. To attempt to develop improved curricular materials, a workshop was organized that combined the teaching expertise of a number of secondary school teachers with the research experience of a number of scientific experts to produce lesson plans centered around trace metal cycling and toxicity. These areas are important current research topics but are often neglected in pre-college education. A significant portion of the material presented at the workshop involved the use of visual and hands-on demonstrations of chemical principles that relate to geochemical processes that impact the environment. Materials provided to the instructors included suggestions for hands-on, interactive activities for use in the classroom, as well as teacher-administered demonstrations that would explore the physical and chemical bases of the principles being discussed. The participants all expressed positive feedback in terms of their confidence to administer student-centered lesson plans on the topic of biogeochemistry and their personal understanding of the subject matter, and a number of lesson plans were developed for use by the teachers to integrate the scientific principles discussed in the body of their courses.

Most subunits of the alphabeta deltaepsilon gammaepsilon zetazeta T cell antigen receptor (TCR) complex associate with the molecular chaperone calnexin shortly after their synthesis in the endoplasmic reticulum, including clonotypic TCRalpha,beta molecules and invariant CD3gamma,delta,epsilon chains. While calnexin interaction is suggested to be important for the stability of newly synthesized TCRalpha subunits, the role of calnexin in the survival and assembly of remaining TCR components is unknown. Here we evaluated the expression of TCR proteins in CEM T cells and the calnexin-deficient CEM variant CEM.NK(R). We found that CEM and CEM.NK(R) cells constitutively synthesized all TCR subunits except for TCRalpha and that CD3gamma,delta,epsilon components and CD3-beta complexes were effectively assembled together in both cell types. The stability and folding of core CD3epsilon chains were similar in CEM and CEM.NK(R) cells. Interestingly, TCRalpha synthesis was differentially induced by phorbol myristate acetate treatment in CEM and CEM.NK(R) cells and TCRalpha proteins synthesized in CEM.NK(R) cells showed reduced survival compared to those made in CEM cells. Importantly, these data show that TCR complexes were inducibly expressed on CEM.NK(R) cells in the absence of calnexin synthesis. These results demonstrate that TCR complexes can be expressed in the absence of calnexin and suggest that the role of calnexin in the quality control of TCR assembly is primarily restricted to the stabilization of newly synthesized TCRalpha proteins.

Most T lymphocytes express on their surfaces a multisubunit receptor complex, the T cell antigen receptor (TCR) containing alpha, beta, gamma, delta, epsilon, and zeta molecules, that has been widely studied as a model system for protein quality control. Although the parameters of TCR assembly are relatively well established, little information exists regarding the stage(s) of TCR oligomerization where folding of TCR proteins is completed. Here we evaluated the modification of TCR glycoproteins by the endoplasmic reticulum folding sensor enzyme UDP-glucose:glycoprotein glucosyltransferase (GT) as a unique and sensitive indicator of how TCR subunits assembled into multisubunit complexes are perceived by the endoplasmic reticulum quality control system. These results demonstrate that all TCR subunits containing N-glycans were modified by GT and that TCR proteins were differentially reglucosylated during their assembly with partner TCR chains. Importantly, these data show that GT modification of most TCR subunits persisted until assembly of CD3alpha beta chains and formation of CD3-associated, disulfide-linked alpha beta heterodimers. These studies provide a novel evaluation of the folding status of TCR glycoproteins during their assembly into multisubunit complexes and are consistent with the concept that TCR folding is finalized convergent with formation of alpha beta delta epsilon gamma epsilon complexes.

Little is known about the microbial communities carried in wind-eroded sediments from various soil types and land management systems. The novel technique of pyrosequencing promises to expand our understanding of the microbial diversity of soils and eroded sediments because it can sequence 10 to 100 times more DNA fragments than previous techniques, providing enhanced exploration into what microbes are being lost from soil due to wind erosion. Our study evaluated the bacterial diversity of two types of wind-eroded sediments collected from three different organic-rich soils in Michigan using a portable field wind tunnel. The wind-eroded sediments evaluated were a coarse sized fraction with 66% of particles >106 孠(coarse eroded sediment) and a finer eroded sediment with 72% of particles

Evaluation of the soil rhizosphere has been limited by the lack of robust assessments that can explore the vast complex structure and diversity of soil microbial communities. Our objective was to combine fatty acid methyl ester (FAME) and pyrosequencing techniques to evaluate soil microbial community structure and diversity. In addition, we evaluated biogeochemical functionality of the microbial communities via enzymatic activities of nutrient cycling. Samples were taken from a silt loam at 0–10 and 10–20 cm in an organic farm under lettuce (Lactuca sativa), potato (Solanum tuberosum), onion (Allium cepa L), broccoli (Brassica oleracea var. botrytis) and Tall fescue pasture grass (Festuca arundinacea). Several FAMEs (a15:0, i15:0, i15:1, i16:0, a17:0, i17:0, 10Me17:0, cy17:0, 16:1ω5c and 18:1ω9c) varied among the crop rhizospheres. FAME profiles of the soil microbial community under pasture showed a higher fungal:bacterial ratio compared to the soil under lettuce, potato, onion, and broccoli. Soil under potato showed higher sum of fungal FAME indicators compared to broccoli, onion and lettuce. Microbial biomass C and enzyme activities associated with pasture and potato were higher than the other rhizospheres. The lowest soil microbial biomass C and enzyme activities were found under onion. Pyrosequencing revealed significant differences regarding the maximum operational taxonomic units (OTU) at 3% dissimilarity level (roughly corresponding to the bacterial species level) at 0–10 cm (581.7–770.0) compared to 10–20 cm (563.3–727.7) soil depths. The lowest OTUs detected at 0–10 cm were under broccoli (581.7); whereas the lowest OTUs found at 10–20 cm were under potato (563.3). The predominant phyla (85%) in this soil at both depths were Bacteroidetes (i.e., Flavobacteria, Sphingobacteria), and Proteobacteria. Flavobacteriaceae and Xanthomonadaceae were predominant under broccoli. Rhizobiaceae, Hyphomicrobiaceae, and Acidobacteriaceae were more abundant under pasture compared to the cultivated soils under broccoli, potato, onion and lettuce. This study found significant differences in microbial community structure and diversity, and enzyme activities of nutrient cycling in this organic farming system under different rhizospheres, which can have implications in soil health and metabolic functioning, and the yield and nutritional value of each crop.

There is still a lack of understanding of how soil microbial community distribution is controlled by wind erosion. This information is of international concern as eroded sediments can potentially carry away the active labile organic soil particulates containing key microorganisms involved in soil biogeochemical processes, which can have a negative impact on the quality and functional potential of the soil. Pyrosequencing techniques promises to expand our understanding of the vast microbial diversity with respect to soils that experience high rates of wind erosion; because it is able to sequence 10-100 times more DNA fragments than previous techniques (traditional cloning). Our study evaluated the bacterial diversity on coarse and fine dust collected from three different silty soils in Michigan by using a portable field wind tunnel instrument. Our results indicated that Acidobacter was the predominant bacteria in these soils as well as the predominant bacteria carried via wind dispersion in coarse and fine dust from these soil sources. Soil 1, which had higher P levels than the other 2 soils, pH was basically 6 and it had higher organic matter (OM) content (47.3-55%), while showing this order of bacterial predominance: Acidobacter, Streptomyces, Levilinea, Patulibacter and Gemmatimonas. Although Streptomyces was the second most abundant bacteria in soil source 1, fine dust did not carry this species, and Levilinea was the second most predominant bacteria in this dust. Soil 2, which had lower P levels than soil 1 (within a range of 122-136 mg P kg-1), pH of about 5.5, and an intermediate OM content (42.9%) also showed the species predominance of Acidobacter followed by Patulibacter, Conexibacter, Rhizobium and Levilinea. Three of the 5 predominant bacteria in the soil source were also predominant in the fine dust except for Conexibacter and Rhizobium. Soil 3 had the lowest OM content (16.3-20.8%) of the 3 soils evaluated, and it had an average pH of 5.7, and P levels within a range of 123-153 P mg kg-1. This soil also showed a predominance of Acidobacter followed by Patulibacter, Rhizobium, Gemmatimonas, and Conexibacter. In addition to Acidobacter in fine dust, Conexibacter and Patulibacter were also carried. The coarse dust samples collected from these 3 soils demonstrated some differences in bacterial distribution compared to the fine dust, which may indicate that fine dust dispersion caused by wind erosion is the major carrier of soil predominant bacteria. The highest abundance of Acidobacter is explained by the acidic pH of these soils, and thus, it appears that they play an important ecological role in these soils functioning. Our findings suggested that bacteria carried in coarse or fine dust represent fingerprints of the soil source, but certain specific group of bacteria was more abundant in fine dust than coarse dust, revealing different niches in these soils. We are in the process of identifying the ecological role of the bacterial groups carried in these dust samples as they can have important implications on the soil sustainability and functioning. This research is focused on coupling the diversity of the soil microbial communities carried by wind erosion with biogeochemical functionality using enzymatic activities involved in nutrient cycling. This will allow us to identify keystone microbial species-assemblages associated with biogeochemical processes of the soil source.

Correct assembly of all TCR complex polypeptides is essential for its cell surface expression and function. The transmembrane region of the TCRalpha chain is highly conserved and to gain insight into the structural and functional role of these residues, single amino acid substitutions were introduced and surface expression and signaling ability studied in T hybridoma cells. Introduction of acid residues within the TCRalpha chain transmembrane region were mostly tolerated, indicating that the net charge within this region of the TCR complex is not crucial to either assembly or signaling. However, mutations of leucine 112 or phenylalanine 127 to aspartic acids (L112D or F127D, respectively) resulted in dramatic loss of surface expression and, therefore, their signaling ability. Intracellular flow cytometry showed that the mutant TCRalpha polypeptides were present at levels comparable to wild-type, indicating that the reduced surface expression was not a consequence of impaired protein survival. The defect was characterized by immunoprecipitation and showed that residues L112 and F127 were involved in early interactions with the CD3 complex. A large proportion of the TCRalpha chain mutants L112D and F127D consisted of immature protein, indicative of a problem during early assembly of the TCR. Our findings provide evidence for the involvement of the conserved L112 and F127 residues of the TCRalpha chain transmembrane region in the assembly process of the TCR complex.