Climate change and C4 and C3 grasses in a midlatitude dryland steppe.

Climate change is projected to alter the structure of plant communities due to increasing temperatures and changes to precipitation patterns, particularly in midlatitude dryland ecosystems. Modifications to climatic suitability may lead to major community changes such as altered dominant plant functional types. Previous studies have indicated that climatic suitability is likely to increase for C4 grasses and decrease for C3 grasses in the Western United States. However, if no C4 grass species currently exist to serve as a propagule source, expansion into areas of increased suitability will be limited. We conducted a field and modeling study in the Upper Green River Basin (UGRB) of Western Wyoming to determine if (1) C4 grasses are present to provide a propagule source and (2) C4 grasses are likely to increase in importance relative to C3 grasses due to climatic changes. We searched 44 sites for C4 grasses to establish presence, and modeled suitability at 35 sites using 17 Global Climate Models, two greenhouse gas Representative Concentration Pathways (RCPs; 4.5 and 8.5), and two time periods (mid- and late century; 2030-2060 and 2070-2099, respectively). We found C4 grasses at 10 of the 44 sites, indicating that there is a present propagule source. Our model projected increases in suitability for both C3 and C4 grasses across sites for all RCPs and time periods. In the mid-century RCP 4.5 scenario, the C3 functional type increased in projected biomass in 29 of 35 sites, and the C4 type increased in 31 sites. In this scenario, C3 grasses increased in projected biomass by a median 4 g m-2 (5% change), and C4 grass biomass increased by a median 8 g m-2 (21% change). Our study suggests that climate change will increase climatic suitability for grasses across the UGRB, and that all requirements are in place for C4 grasses to increase in abundance.

Power Bars: Mormon Crickets Get Immunity Boost from Eating Grasshoppers.

In addition to feeding on plants, Mormon crickets Anabrus simplex Haldeman, 1852 predate on invertebrates, including one another, which effectively drives their migration. Carnivory derives from lack of dietary protein, with Mormon crickets deprived of protein having less phenoloxidase (PO) available to combat foreign invaders, such as fungal pathogens. Because Mormon crickets commonly occur with grasshoppers that feed on the same plants, we investigated interactions between grasshoppers and Mormon crickets, and hypothesized that if Mormon crickets are predatory on grasshoppers, grasshopper abundance would influence the protein available to Mormon crickets and their immunity. In a field setting, we varied densities of Mormon crickets (0, 10, or 20 per cage) and grasshoppers Melanoplus borealis (0, 15, 30, or 45) in 68 1-m2 cages. After one month, we measured Mormon cricket dietary preferences and PO activity. As predicted, artificial diet consumption shifted away from protein as grasshopper density increased, and immunocompetence, as measured by PO activity, also increased with grasshopper availability. Although nitrogen availability in the vegetation decreased with increasing insect density, predation became an important source of protein for Mormon crickets that enhanced immunity. Grasshoppers can be an important source of dietary protein for Mormon crickets, with prey availability affecting Mormon cricket immunity to diseases.

Protogygiapryorensis Crabo & Kirst (Insecta, Lepidoptera, Noctuidae, Noctuinae, Noctuini), a new moth species from Montana, United States of America.

Protogygiapryorensissp. nov. (Insecta, Lepidoptera, Noctuidae, Noctuinae, Noctuini) is described from a single arid active sands habitat in the south foothills of the Pryor Mountains in south-central Montana, USA. It flies during early May. The adult male and its genitalia are illustrated and are compared to similar Protogygia McDunnough species. The female is unknown. Protogygia species groups are discussed and P.pryorensis is assigned to the album-group. The Pryor Mountains foothill habitat of P.pryorensis is described and illustrated.

Using Opportunistic Samples to Monitor West Nile Virus Infection Status in Greater Sage-grouse (Centrocercus urophasianus) in Wyoming, USA (2020-22).

The frequency of arthropod-borne viral disease in naïve hosts is subject to change based on complex interactions among vector, host, virus, and external factors (e.g., climate). Thus, continual monitoring for both disease susceptibility and host infection dynamics is needed, especially for viruses that have proven detrimental to the health of wildlife hosts of conservation concern. The Greater Sage-grouse (Centrocercus urophasianus) is a gamebird of ecological and economic importance in the western United States for which population declines have led to a Near Threatened conservation status. Although these declines have mainly been attributed to habitat loss, West Nile virus (WNV) also poses a threat, with regional transmission potentially fueled by anthropogenic landscape alterations that may facilitate mosquito breeding. With limited WNV monitoring in Greater Sage-grouse after recognition of high susceptibility to mortality early after its initial detection in the western United States, the potential long-term impacts of WNV on this species are poorly understood. We used the plaque reduction neutralization test of filter paper strip-eluted sera to assess for anti-WNV antibodies, indicating prior infection, in opportunistically collected samples. From 2020 to 2022, 85 Greater Sage-grouse in Wyoming were sampled; none had anti-WNV antibodies. This result corroborates findings of previous studies documenting low seroprevalence. With the tenuous conservation status of the species, all potential population health risks should be considered in future management strategies, especially in the face of changing climate and landscapes.

Mortality in Wild Turkeys (Meleagris gallopavo) Associated with Natural Infection with H5N1 Highly Pathogenic Avian Influenza Virus (HPAIV) Subclade 2.3.4.4.

A Eurasian strain of H5N1 highly pathogenic avian influenza virus (HPAIV) was first detected in North America in December 2021 and has since been confirmed in numerous wild and domestic avian species. In April 2022, 41 Wild Turkeys (Meleagris gallopavo) were found dead in Johnson County, Wyoming, USA adjacent to a property with confirmed HPAIV in a backyard poultry flock. Oropharyngeal swabs were collected from 11 of the 41 turkeys and necropsy was performed on seven. Avian influenza virus RNA was detected in all 11 turkeys by real-time reverse-transcription PCR. Acute, multiorgan necrosis was observed grossly and identified in all seven turkeys evaluated by histopathology, most consistently in the lung, spleen, liver, gastrointestinal tract, and gonads. Lesions indicate high virulence of subclade 2.3.4.4b H5N1 HPAIV in Wild Turkeys, with infections presenting as clusters of acute mortality. Although documented cases of HPAIV in Wild Turkeys are rare, these findings signify a risk of spillback from domestic poultry, which may be heightened by the recent rise in backyard poultry ownership and the use of peridomestic habitat by wild birds. Additional research is needed to better understand the risk of disease transmission at the interface of Wild Turkeys and backyard poultry and the potential conservation and management implications of HPAIV in wild gallinaceous birds.

Opioid-Involved Overdose Vulnerability in Wyoming: Measuring Risk in a Rural Environment.

BACKGROUND: Between 2009 and 2019 opioid-involved fatal overdose rates increased by 45% and the average opioid dispensing rate in Wyoming was higher than the national average. The opioid crisis is shaped by a complex set of socioeconomic, geopolitical, and health-related variables. We conducted a vulnerability assessment to identify Wyoming counties at higher risk of opioid-related harm, factors associated with this risk, and areas in need of overdose treatment access to inform priority responses. METHODS: We compiled 2016 to 2018 county-level aggregated and de-identified data. We created risk maps and ran spatial analyses in a geographic information system to depict the spatial distribution of overdose-related measures. We used addresses of opioid treatment programs and buprenorphine providers to develop drive-time maps and ran 2-step floating catchment area analyses to measure accessibility to treatment. We used a straightforward and replicable weighted ranks approach to calculate final county vulnerability scores and rankings from most to least vulnerable. FINDINGS: We found Hot Springs, Carbon, Natrona, Fremont, and Sweetwater Counties to be most vulnerable to opioid-involved overdose fatalities. Opioid prescribing rates were highest in Hot Springs County (97 per 100 persons), almost two times the national average (51 per 100 persons). Statewide, there were over 90 buprenorphine-waivered providers, however accessibility to these clinicians was limited to urban centers. Most individuals lived further than a four-hour round-trip drive to the nearest methadone treatment program. CONCLUSIONS: Identifying Wyoming counties with high opioid overdose vulnerabilities and limited access to overdose treatment can inform public health and harm reduction responses.

Policy-Driven Potential for Deploying Carbon Capture and Sequestration in a Fossil-Rich Power Sector.

In 2020, the Wyoming Legislature enacted House Bill No. 0200 (HB0200), which requires utilities to generate a percentage of dispatchable and reliable low-carbon electricity by 2030. This state requirement must take into consideration "any potentially expiring federal tax credits", such as the federal Section 45Q tax credit. This study aims to examine the potential role of economic and policy incentives that facilitate carbon capture and sequestration (CCS) deployment. A unit-level retrofit analysis shows that deploying CCS at existing coal-fired power plants in Wyoming to meet the HB0200 emission limit would decrease the net efficiency by 29% and increase the levelized cost of electricity by 237% on the fleet average. The CO2 avoidance cost varies by unit from $65/t to 201/t, which reveals economic challenges for CCS retrofits. However, the current tax credit of $50 per metric ton of CO2 for saline-reservoir storage can lower the avoidance cost by 47% on the fleet average. The proposed enhancement of the tax credit to $85/t would offset the added cost for CCS deployment for a total capacity of 3.4 GW. Joint policy and economic incentives can encourage fossil fuel abatement to play a firm role in energy transition.

A novel camera trapping method for individually identifying pumas by facial features.

Camera traps (CTs), used in conjunction with capture-mark-recapture analyses (CMR; photo-CMR), are a valuable tool for estimating abundances of rare and elusive wildlife. However, a critical requirement of photo-CMR is that individuals are identifiable in CT images (photo-ID). Thus, photo-CMR is generally limited to species with conspicuous pelage patterns (e.g., stripes or spots) using lateral-view images from CTs stationed along travel paths. Pumas (Puma concolor) are an elusive species for which CTs are highly effective at collecting image data, but their suitability to photo-ID is controversial due to their lack of pelage markings. For a wide range of taxa, facial features are useful for photo-ID, but this method has generally been limited to images collected with traditional handheld cameras. Here, we evaluate the feasibility of using puma facial features for photo-ID in a CT framework. We consider two issues: (1) the ability to capture puma facial images using CTs, and (2) whether facial images improve human ability to photo-ID pumas. We tested a novel CT accessory that used light and sound to attract the attention of pumas, thereby collecting face images for use in photo-ID. Face captures rates increased at CTs that included the accessory (n = 208, χ 2 = 43.23, p ≤ .001). To evaluate if puma faces improve photo-ID, we measured the inter-rater agreement of 5 independent assessments of photo-ID for 16 of our puma face capture events. Agreement was moderate to good (Fleiss' kappa = 0.54, 95% CI = 0.48-0.60), and was 92.90% greater than a previously published kappa using conventional CT methods. This study is the first time that such a technique has been used for photo-ID, and we believe a promising demonstration of how photo-ID may be feasible for an elusive but unmarked species.

A NEW SPECIES OF EIMERIA (APICOMPLEXA: EIMERIIDAE) FROM THE OLIVE-BACKED POCKET MOUSE, PEROGNATHUS FASCIATUS (RODENTIA: HETEROMYIDAE: PEROGNATHINAE), FROM WYOMING.

Forty-nine olive-backed pocket mice, Perognathus fasciatus were collected during 2011 and 2012 from 4 sites in Wyoming and examined for coccidian parasites. Fifteen (31%) were found to be passing oocysts of a new species of Eimeria Schneider, 1875. Sporulated oocysts of Eimeria fasciata n. sp. are ellipsoidal to ovoidal, 23.3 × 20.7 (19-27 × 17-25) µm, with a shape index of 1.1; they typically contain a single, smooth, bubble-like oocyst residuum. Oocysts possess 1-2 polar granules, lack a micropyle, and are bilayered with a thickness of 1.3 µm. Sporocysts are ovoidal, 10.0 × 8.2 (8-12 × 7-10) µm, with a shape index of 1.2; they contain a sporocyst residuum that appears similar to a cluster of 1-8 grapes. The Stieda body is small, appearing flattened to knobby, and there are no subStieda or paraStieda bodies. This new eimerian represents the only coccidian, to date, reported from P. fasciatus, as well as the only species from any heteromyid rodent in Wyoming.

Survey of coyotes, red foxes and wolves from Wyoming, USA, for Echinococcus granulosus s. l.

The paraphyletic group Echinococcus granulosus sensu lato is comprised of parasitic tapeworms of wild and domestic canids such as wolves (Canis lupus) and coyotes (Canis latrans), which serve as definitive hosts, and ungulates, which are the intermediate hosts. Members of this tapeworm group are characterized by both cosmopolitan distribution and zoonotic disease potential. This survey (conducted from 2012 through 2017) was designed to provide insight into the prevalence and distribution of this parasite in wild canids in Wyoming. Echinococcus sp. infections were documented in 14 of 22 gray wolves (63.6%), 1 of 182 coyotes (0.55%) and 0 of 5 red foxes (Vulpes fulva). Echinococcus granulosus s. l. was confirmed in 4 of these 14 specimens obtained from wolves with two parasite specimens corresponding morphologically with E. canadensis (G8/G10). These results suggest that wolves serve as the major definitive host of E. granulosus s. l. in Wyoming, while coyotes do not play an equivalent role. Limited sample size precludes evaluation of the importance of the red fox as a favorable definitive host. Whereas this study documents the occurrence of E. granulosus s. l. in Wyoming, the zoonotic disease risk does not appear to be high. Education remains the key to disease prevention, coupled with good hygienic practices by humans and anthelmintic treatment of domestic dogs exhibiting elevated risk of exposure.

The osteology and phylogenetic position of the loricatan (Archosauria: Pseudosuchia) Heptasuchus clarki, from the ?Mid-Upper Triassic, southeastern Big Horn Mountains, Central Wyoming (USA).

Loricatan pseudosuchians (known as "rauisuchians") typically consist of poorly understood fragmentary remains known worldwide from the Middle Triassic to the end of the Triassic Period. Renewed interest and the discovery of more complete specimens recently revolutionized our understanding of the relationships of archosaurs, the origin of Crocodylomorpha, and the paleobiology of these animals. However, there are still few loricatans known from the Middle to early portion of the Late Triassic and the forms that occur during this time are largely known from southern Pangea or Europe. Heptasuchus clarki was the first formally recognized North American "rauisuchian" and was collected from a poorly sampled and disparately fossiliferous sequence of Triassic strata in North America. Exposed along the trend of the Casper Arch flanking the southeastern Big Horn Mountains, the type locality of Heptasuchus clarki occurs within a sequence of red beds above the Alcova Limestone and Crow Mountain formations within the Chugwater Group. The age of the type locality is poorly constrained to the Middle-early Late Triassic and is likely similar to or just older than that of the Popo Agie Formation assemblage from the western portion of Wyoming. The holotype consists of associated cranial elements found in situ, and the referred specimens consist of crania and postcrania. Thus, about 30% of the osteology of the taxon is preserved. All of the pseudosuchian elements collected at the locality appear to belong to Heptasuchus clarki and the taxon is not a chimera as previously hypothesized. Heptasuchus clarki is distinct from all other archosaurs by the presence of large, posteriorly directed flanges on the parabasisphenoid and a distinct, orbit-overhanging postfrontal. Our phylogenetic hypothesis posits a sister-taxon relationship between Heptasuchus clarki and the Ladinian-aged Batrachotomus kupferzellensis from current-day Germany within Loricata. These two taxa share a number of apomorphies from across the skull and their phylogenetic position further supports 'rauisuchian' paraphyly. A minimum of three individuals of Heptasuchus are present at the type locality suggesting that a group of individuals died together, similar to other aggregations of loricatans (e.g., Heptasuchus, Batrachotomus, Decuriasuchus, Postosuchus).

Mycoplasma bovis Infections in Free-Ranging Pronghorn, Wyoming, USA.

Mycoplasma bovis is 1 of several bacterial pathogens associated with pneumonia in cattle. Its role in pneumonia of free-ranging ungulates has not been established. Over a 3-month period in early 2019, ¼60 free-ranging pronghorn with signs of respiratory disease died in northeast Wyoming, USA. A consistent finding in submitted carcasses was severe fibrinosuppurative pleuropneumonia and detection of M. bovis by PCR and immunohistochemical analysis. Multilocus sequence typing of isolates from 4 animals revealed that all have a deletion in 1 of the target genes, adh-1. A retrospective survey by PCR and immunohistochemical analysis of paraffin-embedded lung from 20 pronghorn that died with and without pneumonia during 2007-2018 yielded negative results. These findings indicate that a distinct strain of M. bovis was associated with fatal pneumonia in this group of pronghorn.

Assessment of Drivers' Perceptions of Connected Vehicle-Human Machine Interface for Driving Under Adverse Weather Conditions: Preliminary Findings From Wyoming.

Connected vehicle (CV) technology aims to improve drivers' situational awareness through audible and visual warnings displayed on a human-machine interface (HMI), thus reducing crashes caused by human error. This paper developed a driving simulator test bed to assess the readability and usefulness of the Wyoming CV applications. A total number of 26 professional drivers were recruited to participate in a driving-simulator study. Prior to driving the simulator, the participants were trained on both the concept of CV technology and the developed CV applications as well as the operation of the driving simulator. Three driving simulation scenarios were designed. For each scenario, participants drove two times: one with the HMI turned on and another one with the HMI turned off. After driving the simulator, a comprehensive revealed-preference survey was employed to collect the participants' perceptions of CV technology and Wyoming CV applications. Results show that the Wyoming CV applications were most favored under poor-visibility driving conditions. Among the Wyoming CV applications, forward collision warning and rerouting applications were experienced as the most useful. Approximately 89% of the participants stated that the Wyoming CV applications provided them with improved road condition information and increased their experienced safety while driving; 65% of the participants stated the CV applications and the HMI did not introduce distraction from the primary task of driving. Finally, this paper concludes that the design of CV HMI needs to balance a trade-off between the readability of the warnings and drivers' capability to safely recognize and timely respond to the received warnings.

An Integrated Microsimulation Approach for Safety Performance Assessment of the Wyoming Connected Vehicle Pilot Deployment Program.

The 402-mile of Interstate 80 in Wyoming was selected by the U.S. Department of Transportation to develop, test, and deploy a suite of Connected Vehicle (CV) applications (WYDOT CV Pilot). It is expected that after full deployment of CV technology, the pilot will improve safety and mobility under adverse weather conditions by creating new ways to communicate road and travel information to both drivers and fleet managers. In this regard, this research employed an integrated microsimulation modeling approach to assess the safety performance of the WYDOT CV Pilot. A 23-mile representative I-80 corridor was selected for developing the microsimulation models. Traffic flow and driving behavior data under winter snowy weather condition were collected to calibrate the baseline microsimulation model. A driving simulator experiment was conducted to quantitatively investigate the impacts of CV technology on driving behavior; accordingly, the driving behavior data under CV environment were employed to properly update the calibrated CV microsimulation models. The safety effectiveness of the WYDOT CV Pilot were assessed for various demand levels and CV penetration rates. It was concluded that WYDOT CV applications increased drivers' situation awareness under adverse weather conditions, and thus reduced the crash risk. The reductions in conflicts displayed a decreasing trend with the increase of CV penetration rates, but the reduction was not significant when CV penetration was lower than 10 percent. The maximum reduction in conflicts was 85 percent when all trucks were equipped with CV technology.

Connected vehicle real-time traveler information messages for freeway speed harmonization under adverse weather conditions: Trajectory level analysis using driving simulator.

This paper employed a high-fidelity driving simulator to investigate the impacts of the Wyoming Department of Transportation (WYDOT) Connected Vehicle (CV) Pilot's Traveler Information Messages (TIMs) on drivers' speed selection and the safety benefits of their speed harmonization. Three driving simulator experiment scenarios were developed to simulate the typical traffic and weather conditions on the rural Interstate 80 (I-80) in Wyoming. A total of 25 professional drivers from the WYDOT and trucking industry were recruited to participate in the driving simulator experiment. Participants' instantaneous speeds at various locations were collected to reveal the effects of CV TIMs on their speed selection. The results showed that average speed profiles under CV scenarios were generally lower than under baseline scenarios, particularly for winter conditions (snowy and severe weather). The variance of speed under CV scenarios was found to be significantly lower than the baseline scenarios, indicating that CV TIMs have the potential to harmonize the variations in speed. In addition, for the work zone driving simulator experiment, this research revealed that the mean time-to-collision (TTC) under baseline scenario is approximately 40 % lower than CV scenario, and the mean deceleration to avoid a crash (DRAC) under baseline scenario is approximately 19.3 % higher than CV scenario. These findings suggest that CV TIMs can reduce the risk of crashes. Research findings would provide the WYDOT with early insights into the effectiveness of CV TIMs, which could assist with developing more efficient transportation management strategies under adverse weather conditions.

Drought reshuffles plant phenology and reduces the foraging benefit of green-wave surfing for a migratory ungulate.

To increase resource gain, many herbivores pace their migration with the flush of nutritious plant green-up that progresses across the landscape (termed "green-wave surfing"). Despite concerns about the effects of climate change on migratory species and the critical role of plant phenology in mediating the ability of ungulates to surf, little is known about how drought shapes the green wave and influences the foraging benefits of migration. With a 19 year dataset on drought and plant phenology across 99 unique migratory routes of mule deer (Odocoileus hemionus) in western Wyoming, United States, we show that drought shortened the duration of spring green-up by approximately twofold (2.5 weeks) and resulted in less sequential green-up along migratory routes. We investigated the possibility that some routes were buffered from the effects of drought (i.e., routes that maintained long green-up duration irrespective of drought intensity). We found no evidence of drought-buffered routes. Instead, routes with the longest green-up in non-drought years also were the most affected by drought. Despite phenological changes along the migratory route, mule deer closely followed drought-altered green waves during migration. Migrating deer did not experience a trophic mismatch with the green wave during drought. Instead, the shorter window of green-up caused by drought reduced the opportunity to accumulate forage resources during rapid spring migrations. Our work highlights the synchronization of phenological events as an important mechanism by which climate change can negatively affect migratory species by reducing the temporal availability of key food resources. For migratory herbivores, climate change poses a new and growing threat by altering resource phenology and diminishing the foraging benefit of migration.

First report of the bent seed gall nematode, Anguina agrostis (Steinbuch, 1799) Filipjev, 1936 from Poa palustris L. in Wyoming, USA.

In September 2020, several plants of fowl bluegrass, Poa palustris with seed galls were collected on a bank of river in Teton County, Wyoming, USA. Isolated nematodes were identified by both morphological and molecular methods as Anguina agrostis. This is a first report of A. agrostis in Wyoming and its report on fowl bluegrass.

Investigating the effect of geometric dimensions of median traffic barriers on crashes: Crash analysis of interstate roads in Wyoming using actual crash datasets.

INTRODUCTION: Despite the numerous safety studies done on traffic barriers' performance assessment, the effect of variables such as traffic barrier's height has not been identified considering a comprehensive actual crash data analysis. This study seeks to identify the impact of geometric variables (i.e., height, post-spacing, sideslope ratio, and lateral offset) on median traffic barriers' performance in crashes on interstate roads. METHOD: Geometric dimensions of over 110 miles median traffic barriers on interstate Wyoming roads were inventoried in a field survey between 2016 and 2018. Then, the traffic barrier data collected was combined with historical crash records, traffic volume data, road geometric characteristics, and weather condition data to provide a comprehensive dataset for the analysis. Finally, an ordered logit model with random-parameters was developed for the severity of traffic barrier crashes. Based on the results, traffic barrier's height was found to impact crash severity. RESULTS: Crashes involving cable barriers with a height between 30″ and 42″ were less severe than other traffic barrier types, while concrete barriers with a height shorter than 32″ were more likely involved with severe injury crashes. As another important finding, the post-spacing of 6.1-6.3 ft. was identified as the least severe range in W-beam barriers. PRACTICAL APPLICATIONS: The results show that using flare barriers should reduce the number of crashes compared to parallel barriers.

Validating DNA Extraction Protocols for Bentonite Clay.

Bentonite clay is an integral component of the engineered barrier system of deep geological repositories (DGRs) that are planned for the long-term storage of high-level radioactive waste. Although nucleic acid extraction and analysis can provide powerful qualitative and quantitative data reflecting the presence, abundance, and functional potential of microorganisms within DGR materials, extraction of microbial DNA from bentonite clay is challenging due to the low biomass and adsorption of nucleic acids to the charged clay matrix. In this study, we used quantitative PCR, gel fingerprinting, and high-throughput sequencing of 16S rRNA gene amplicons to assess DNA extraction efficiency from natural MX-80 bentonite and the same material "spiked" with Escherichia coli genomic DNA. Extraction protocols were tested without additives and with casein and phosphate as blocking agents. Although we demonstrate improved DNA recovery by blocking agents at relatively high DNA spiking concentrations, at relatively low spiking concentrations, we detected a high proportion of contaminant nucleic acids from blocking agents that masked sample-specific microbial profile data. Because bacterial genomic DNA associated with casein preparations was insufficiently removed by UV treatment, casein is not recommended as an additive for DNA extractions from low-biomass samples. Instead, we recommend a kit-based extraction protocol for bentonite clay without additional blocking agents, as tested here and validated with multiple MX-80 bentonite samples, ensuring relatively high DNA recoveries with minimal contamination.IMPORTANCE Extraction of microbial DNA from MX-80 bentonite is challenging due to low biomass and adsorption of nucleic acid molecules to the charged clay matrix. Blocking agents improve DNA recovery, but their impact on microbial community profiles from low-biomass samples has not been characterized well. In this study, we evaluated the effect of casein and phosphate as blocking agents for quantitative recovery of nucleic acids from MX-80 bentonite. Our data justify a simplified framework for analyzing microbial community DNA associated with swelling MX-80 bentonite samples within the context of a deep geological repository for used nuclear fuel. This study is among the first to demonstrate successful extraction of DNA from Wyoming MX-80 bentonite.