Soil Properties and Moisture Synergistically Influence Nontuberculous Mycobacterial Prevalence in Natural Environments of Hawai'i.

Nontuberculous mycobacteria (NTM) are opportunistic pathogens that cause chronic pulmonary disease (PD). NTM infections are thought to be acquired from the environment; however, the basal environmental factors that drive and sustain NTM prevalence are not well understood. The highest prevalence of NTM PD cases in the United States is reported from Hawai'i, which is unique in its climate and soil composition, providing an opportunity to investigate the environmental drivers of NTM prevalence. We used microbiological sampling and spatial logistic regression complemented with fine-scale soil mineralogy to model the probability of NTM presence across the natural landscape of Hawai'i. Over 7 years, we collected and microbiologically cultured 771 samples from 422 geographic sites in natural areas across the Hawaiian Islands for the presence of NTM. NTM were detected in 210 of these samples (27%), with Mycobacterium abscessus being the most frequently isolated species. The probability of NTM presence was highest in expansive soils (those that swell with water) with a high water balance (>1-m difference between rainfall and evapotranspiration) and rich in Fe-oxides/hydroxides. We observed a positive association between NTM presence and iron in wet soils, supporting past studies, but no such association in dry soils. High soil-water balance may facilitate underground movement of NTM into the aquifer system, potentially compounded by expansive capabilities allowing crack formation under drought conditions, representing further possible avenues for aquifer infiltration. These results suggest both precipitation and soil properties are mechanisms by which surface NTM may reach the human water supply. IMPORTANCE Nontuberculous mycobacteria (NTM) are ubiquitous in the environment, being found commonly in soils and natural bodies of freshwater. However, little is known about the environmental niches of NTM and how they relate to NTM prevalence in homes and other human-dominated areas. To characterize NTM environmental associations, we collected and cultured 771 samples from 422 geographic sites in natural areas across Hawai'i, the U.S. state with the highest prevalence of NTM pulmonary disease. We show that the environmental niches of NTM are most associated with highly expansive, moist soils containing high levels of iron oxides/hydroxides. Understanding the factors associated with NTM presence in the natural environment will be crucial for identifying potential mechanisms and risk factors associated with NTM infiltration into water supplies, which are ultimately piped into homes where most exposure risk is thought to occur.

Assessment of Soil Features on the Growth of Environmental Nontuberculous Mycobacterial Isolates from Hawai'i.

Environmental nontuberculous mycobacteria (NTM), with the potential to cause opportunistic lung infections, can reside in soil. This might be particularly relevant in Hawai'i, a geographic hot spot for NTM infections and whose soil composition differs from many other areas of the world. Soil components are likely to contribute to NTM prevalence in certain niches as food sources or attachment scaffolds, but the particular types of soils, clays, and minerals that impact NTM growth are not well-defined. Hawai'i soil and chemically weathered rock (saprolite) samples were examined to characterize the microbiome and quantify 11 mineralogical features as well as soil pH. Machine learning methods were applied to identify important soil features influencing the presence of NTM. Next, these features were directly tested in vitro by incubating synthetic clays and minerals in the presence of Mycobacteroides abscessus and Mycobacterium chimaera isolates recovered from the Hawai'i environment, and changes in bacterial growth were determined. Of the components examined, synthetic gibbsite, a mineral form of aluminum hydroxide, inhibited the growth of both M. abscessus and M. chimaera, while other minerals tested showed differential effects on each species. For example, M. abscessus (but not M. chimaera) growth was significantly higher in the presence of hematite, an iron oxide mineral. In contrast, M. chimaera (but not M. abscessus) counts were significantly reduced in the presence of birnessite, a manganese-containing mineral. These studies shed new light on the mineralogic features that promote or inhibit the presence of Hawai'i NTM in Hawai'i soil.IMPORTANCE Globally and in the United States, the prevalence of NTM pulmonary disease-a potentially life-threatening but underdiagnosed chronic illness-is prominently rising. While NTM are ubiquitous in the environment, including in soil, the specific soil components that promote or inhibit NTM growth have not been elucidated. We hypothesized that NTM culture-positive soil contains minerals that promote NTM growth in vitro Because Hawai'i is a hot spot for NTM and a unique geographic archipelago, we examined the composition of Hawai'i soil and identified individual clay, iron, and manganese minerals associated with NTM. Next, individual components were evaluated for their ability to directly modulate NTM growth in culture. In general, gibbsite and some manganese oxides were shown to decrease NTM, whereas iron-containing minerals were associated with higher NTM counts. These data provide new information to guide future analyses of soil-associated factors impacting persistence of these soil bacteria.

Hawaiian Volcanic Ash, an Airborne Fomite for Nontuberculous Mycobacteria.

Nontuberculous mycobacteria (NTM) are environmentally acquired opportunistic pathogens that can cause chronic lung disease. Within the U.S., Hawai'i shows the highest prevalence rates of NTM lung infections. Here, we investigated a potential role for active volcanism at the Kilauea Volcano located on Hawai'i Island in promoting NTM growth and diversity. We recovered NTM that are known to cause lung disease from plumbing biofilms and soils collected from the Kilauea environment. We also discovered viable Mycobacterium avium, Mycobacterium abscessus, and Mycobacterium intracellulare subsp. chimaera on volcanic ash collected during the 2018 Kilauea eruption. Analysis of soil samples showed that NTM prevalence is positively associated with bulk content of phosphorus, sulfur, and total organic carbon. In growth assays, we showed that phosphorus utilization is essential for proliferation of Kilauea-derived NTM, and demonstrate that NTM cultured with volcanic ash adhere to ash surfaces and remain viable. Ambient dust collected on O'ahu concurrent with the 2018 eruption contained abundant fresh volcanic glass, suggestive of inter-island ash transport. Phylogenomic analyses using whole genome sequencing revealed that Kilauea-derived NTM are genetically similar to respiratory isolates identified on other Hawaiian Islands. Consequently, we posit that volcanic eruptions could redistribute environmental microorganisms over large scales. While additional studies are needed to confirm a direct role of ash in NTM dispersal, our results suggest that volcanic particulates harbor and can redistribute NTM and should therefore be studied as a fomite for these burgeoning, environmentally acquired respiratory infections.

Exposure Pathways of Nontuberculous Mycobacteria Through Soil, Streams, and Groundwater, Hawai'i, USA.

Although uncommon, nontuberculous mycobacterial (NTM) pulmonary infection in the Hawaiian Islands has a relatively high incidence and mortality compared to the mainland U.S. As a result, this study examines the possible geological and hydrological pathways by which NTM patients may become infected, including the environmental conditions that may favor growth and transport. Previously suggested infection routes include the inhalation of NTM attached to micro-droplets from infected home plumbing systems and aerosolized dust from garden soil. In this study, we evaluate the possible routes NTM may take from riparian environments, into groundwater, into public water supplies and then into homes. Because NTM are notoriously hydrophobic and prone to attach to surfaces, mineralogy, and surface chemistry of suspended sediment in streams, soils, and rock scrapings suggest that NTM may especially attach to Fe-oxides/hydroxides, and be transported as particles from losing streams to the aquifer on time-scales of minutes to days. Within the aquifer, flow models indicate that water may be drawn into production wells on time scales (months) that permit NTM to survive and enter domestic water supplies. These processes depend on the presence of interconnected fracture networks with sufficient aperture to preclude complete autofiltration. The common occurrence of NTM in and around streams, in addition to wells, implies that the natural and built environments are capable of introducing a source of NTM into domestic water supplies via groundwater withdrawals. This may produce a persistent source of NTM infection to individuals through the presence of NTM-laden biofilms in home plumbing.

Mercury and dissolved organic matter dynamics during snowmelt runoff in a montane watershed, Provo River, Utah, USA.

The mechanisms of Hg and dissolved organic matter (DOM) transport from watersheds to streams remain unclear, especially in snowmelt dominated montane systems. We characterized total Hg concentrations and DOM characteristics during snowmelt by weekly and/or monthly sampling at three locations in the upper Provo River, northern Utah, over two water years (2016 and 2017). Total Hg concentrations increased from <1 ng/L during baseflow to >7 ng/L during the snowmelt period (April-June), with decreasing concentrations from upstream to downstream. Filtered THg concentrations accounted for 65-75% of the unfiltered concentrations, suggesting that Hg is primarily transported in the dissolved phase. Annual THg loading in the upper Provo River was approximately 1 kg/yr, with ~90% of the flux occurring during the snowmelt period. Filtered THg concentrations were strongly correlated with in-situ fluorescence DOM (fDOM) measurements, allowing for the development of high-resolution proxy THg concentrations. Further, DOM characteristics, evaluated using excitation-emission matrices (EEMs) and parallel factor analysis (PARAFAC), identified the dominance of soil humic and fulvic acid fractions of DOM during runoff. Total Hg concentrations were low in snowpack but elevated in ephemeral streams during snowmelt runoff, indicating that Hg originated from shallow soil water rather than snow. Concentration-discharge relationships revealed clockwise hysteresis patterns, suggesting that Hg was flushed from soils on the rising limb of the hydrograph. Our results demonstrate that a majority of Hg transport occurs with a flux of DOM during the short snowmelt season as shallow soils are flushed by meltwater. The snowmelt-driven Hg pulse is a substantial source to downstream reservoirs and potentially contributes to a fish consumption advisory.

Sediment potentially controls in-lake phosphorus cycling and harmful cyanobacteria in shallow, eutrophic Utah Lake.

Lakes worldwide are impacted by eutrophication and harmful algal or cyanobacteria blooms (HABs) due to excessive nutrients, including legacy P released from sediments in shallow lakes. Utah Lake (northern Utah, USA) is a shallow lake with urban development primarily on the east side of the watershed, providing an opportunity to evaluate HABs in relation to a gradient of legacy sediment P. In this study, we investigated sediment composition and P concentrations in sediment, pore water, and the water column in relation to blooms of harmful cyanobacteria species. Sediments on the east side of the lake had P concentrations up to 1710 mg/kg, corresponding to elevated P concentrations in pore water (up to 10.8 mg/L) and overlying water column (up to 1.7 mg/L). Sediment P concentrations were positively correlated with Fe2O3, CaO, and organic matter abundance, and inversely correlated with SiO2, demonstrating the importance of sediment composition for P sorption and mineral precipitation. Although the sediment contained <3% Fe2O3 by weight, approximately half of the sediment P was associated with redox-sensitive Fe oxide/hydroxide minerals that could be released to the water column under reducing conditions. Cyanobacteria cell counts indicate that blooms of Aphanizomenon flos-aquae and Dolichospermum flosaquae species tend to occur on the east side of Utah Lake, corresponding to areas with elevated P concentrations in the sediment, pore water, and water column. Our findings suggest that shallow lake eutrophication may be a function of P in legacy sediments that contribute to observed HABs in specific locations of shallow lakes.

An objective means of diagnosing anorexia nervosa and bulimia nervosa using 15N/14N and 13C/12C ratios in hair.

An objective means based on the carbon and nitrogen stable isotope analysis of five hairs per individual is presented for distinguishing between individuals with anorexia nervosa and/or bulimia nervosa from non-clinical individuals (i.e. clinically normal controls). Using discriminant analysis, an algorithm has been developed that provides both sensitivity and specificity of 80% in making diagnoses of individuals with these eating disorders. With further refinements, the results suggest that it may be also possible to distinguish between individuals with anorexia or bulimia. Finally, the study shows the value of conducting blind tests and using larger sample sizes of both control and treatment groups. Both groups are needed to validate the diagnostic value of a method and to provide measures of sensitivity and specificity of any diagnostic test.