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The potential association of milk with childhood obesity has been widely debated and researched. Milk is known to contain many bioactive compounds as well as bovine exosomes rich in micro-RNA (miR) that can have effects on various cells, including stem cells. Among them, adipose stem cells (ASC) are particularly interesting due to their role in adipose tissue growth and, thus, obesity. The objective of this study was to evaluate the effect of milk consumption on miR present in circulating exosomes and the transcriptome of ASC in piglets. Piglets were supplemented for 11 weeks with 750 mL of whole milk (n = 6; M) or an isocaloric maltodextrin solution (n = 6; C). After euthanasia, ASC were isolated, quantified, and characterized. RNA was extracted from passage 1 ASC and sequenced. Exosomes were isolated and quantified from the milk and plasma of the pigs at 6-8 hours after milk consumption, and miRs were isolated from exosomes and sequenced. The transfer of exosomes from milk to porcine plasma was assessed by measuring bovine milk-specific miRs and mRNA in exosomes isolated from the plasma of 3 piglets during the first 6h after milk consumption. We observed a higher proportion of exosomes in the 80 nM diameter, enriched in milk, in M vs. C pigs. Over 500 genes were differentially expressed (DEG) in ASC isolated from M vs. C pigs. Bioinformatic analysis of DEG indicated an inhibition of the immune, neuronal, and endocrine systems and insulin-related pathways in ASC of milk-fed pigs compared with maltodextrin-fed pigs. Of the 900 identified miRs in porcine plasma exosomes, only 3 miRs were differentially abundant between the two groups and could target genes associated with neuronal functions. We could not detect exosomal miRs or mRNA transfer from milk to porcine-circulating plasma exosomes. Our data highlights the significant nutrigenomic role of milk consumption on ASC, a finding that does not appear to be attributed to miRs in bovine milk exosomes. The downregulation of insulin resistance and inflammatory-related pathways in the ASC of milk-fed pigs should be further explored in relation to milk and human health. In conclusion, the bioinformatic analyses and the absence of bovine exosomal miRs in porcine plasma suggest that miRs are not vertically transferred from milk exosomes., Competing Interests: The authors have declared that no competing interests exist, (Copyright: © 2024 Swanson et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Presence-absence surveys are frequently used to monitor populations of rare and elusive species. Such data may also be used as a proxy for breeding activity, but links between presence-absence data and higher-order processes must be validated to determine their reliability. The Marbled Murrelet (Brachyramphus marmoratus) is a threatened seabird that nests in older-aged forests along the Pacific Coast. Its nests are exceptionally difficult to find, so we tested whether presence-absence surveys can help identify nesting sites. Between 2018 and 2022 we located 17 trees containing active murrelet nests in the Oregon Coast Range (USA) and 38 trees that purportedly contained no active nests (26 in occupied murrelet stands and 12 in unoccupied stands). Observers surveyed within 200 m of focal trees using standard presence-absence surveys, and we modeled the effects of site status (active nest or control) and distance from the focal tree on probability of recording murrelets. We never detected murrelets in unoccupied control sites. We found some evidence that the probability of recording presence was higher at active nesting sites (0.81, 95% CI: 0.71, 0.88) than at occupied control sites (0.71, 95% CI: 0.64, 0.78) although a null model had similar support. The probability of recording murrelet breeding behaviors in nesting and occupied control sites was 0.20 (95% CI: 0.14, 0.27) regardless of distance to a known active nest. These results suggest that presence-absence surveys may be useful for identifying plausible murrelet nesting habitat, but they are ineffective for identifying active nesting sites. Moreover, we estimated that 20 repeated surveys at a point in space are required to reasonably conclude there are no active nesting sites within 200 m. These findings serve as an important reminder of the limitations that can come with relying on presence-absence data alone to identify breeding sites., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Social interaction and solitude entail tradeoffs. Communicate Bond Belong (CBB) theory holds that social interaction can foster relatedness with others at the cost of social energy, whereas solitude can restore social energy at the cost of relatedness. The current study empirically tests this tradeoff of solitude and its implications for well-being by investigating different degrees of solitude. Less complete degrees of solitude (e.g., no interaction with others) were associated with more relatedness and restoration than more complete degrees of solitude (e.g., no interaction, no potential for it, and no engagement with media), speaking against a tradeoff. Solitude was less detrimental for well-being among individuals who perceived it to be associated with higher restoration and relatedness. Yet, this finding was independent of social energy expenditure, challenging CBB theory. Future work should consider motivations for solitude and longitudinal approaches., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Ross, Campbell. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

DARVO (Deny, Attack, Reverse Victim and Offender) is a response frequently exhibited by perpetrators of wrongdoing after being confronted or held accountable for their harmful behaviors. Consistent with the original conceptualization of DARVO as a strategy used by sex offenders to deflect blame and responsibility, sexual violence survivors report experiencing DARVO from their perpetrators following an assault. The purpose of the current study was to extend research on the connections between DARVO and sexual violence. We examined whether people who use DARVO as a means of responding to confrontations involving a range of wrongdoings also engage in behaviors and ascribe to beliefs that contribute to sexual violence. A sample of 602 university students was recruited to test hypotheses predicting positive associations between individuals' use of DARVO responses, sexual harassment perpetration, and acceptance of rape myths. Supporting predictions, small but positive correlations emerged between study variables. Data from a second sample of 335 community adults from MTurk were analyzed to replicate findings from the undergraduate sample. Results from the community sample also revealed significant associations between DARVO use, sexual harassment perpetration, and rape myth acceptance. Findings offer further confirmation of a link between DARVO and sexual violence and suggest this defensive response is part of a larger worldview that justifies participation in sexual violence and blames victims., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: Jennifer J. Freyd, PhD, is the Founder and President of the Center for Institutional Courage and Professor Emerit of Psychology at the University of Oregon. She is also a Member of the Advisory Committee, 2019-2023, for the Action Collaborative on Preventing Sexual Harassment in Higher Education, National Academies of Science, Engineering, and Medicine. Freyd is the author of the Harvard Press book Betrayal Trauma: The Logic of Forgetting Childhood Abuse. Her most recent book Blind to Betrayal, co-authored with Pamela J. Birrell, was published by John Wiley. For these books Freyd receives royalties. She is often paid honoraria for presentations and she has served as an expert witness on some legal cases for both profit and nonprofit law firms and she has consulted for some governmental and nonprofit organizations. This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2024 Harsey et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The ability to efficiently target loci in the Chlamydia trachomatis genome for deletion remains a desirable goal in the field and new strategies need to be developed and refined. Here we describe the development and application of a lambda red recombineering system for Chlamydia. Using a non-replicative plasmid encoding key lambda Red components and targeting sequences, we demonstrate the efficient deletion of numerous gene targets in the model chlamydial strains C. trachomatis L2/434 and C. muridarum. For initial development of the system, we targeted the incA gene of C. trachomatis for deletion. Deletion mutants containing a chloramphenicol resistance marker were recovered within 3 rounds of selection, or 2 rounds of passaging, in McCoy cells and the resulting clones (CTΔincA) were verified by PCR-based genotyping and whole genome sequencing. Phenotypic assessment of host cells infected with CTΔincA was performed by immunofluorescence microscopy and confirmed the lack of IncA expression and the uniform presence of nonfusogenic vacuoles (inclusions) across CTΔincA-infected monolayers. To explore the utility of this system, we deleted 5 additional candidate virulence factors in C. trachomatis and C. muridarum, including deletions of single and multiple genes. We expect lambda Red recombineering to offer a powerful new strategy for making gene deletion and/or replacement mutants in Chlamydia., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Wang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objective: The epidemiology of COVID-19 has substantially changed since its emergence given the availability of effective vaccines, circulation of different viral variants, and re-infections. We aimed to develop models to predict 30-day COVID-19 hospitalization and death in the Omicron era for contemporary clinical and research applications., Methods: We used comprehensive electronic health records from a national cohort of patients in the Veterans Health Administration (VHA) who tested positive for SARS-CoV-2 between March 1, 2022, and March 31, 2023. Full models incorporated 84 predictors, including demographics, comorbidities, and receipt of COVID-19 vaccinations and anti-SARS-CoV-2 treatments. Parsimonious models included 19 predictors. We created models for 30-day hospitalization or death, 30-day hospitalization, and 30-day all-cause mortality. We used the Super Learner ensemble machine learning algorithm to fit prediction models. Model performance was assessed with the area under the receiver operating characteristic curve (AUC), Brier scores, and calibration intercepts and slopes in a 20% holdout dataset., Results: Models were trained and tested on 198,174 patients, of whom 8% were hospitalized or died within 30 days of testing positive. AUCs for the full models ranged from 0.80 (hospitalization) to 0.91 (death). Brier scores were close to 0, with the lowest error in the mortality model (Brier score: 0.01). All three models were well calibrated with calibration intercepts <0.23 and slopes <1.05. Parsimonious models performed comparably to full models., Conclusions: We developed prediction models that accurately estimate COVID-19 hospitalization and mortality risk following emergence of the Omicron variant and in the setting of COVID-19 vaccinations and antiviral treatments. These models may be used for risk stratification to inform COVID-19 treatment and to identify high-risk patients for inclusion in clinical trials., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Analysis of trapped honey bee pollen for pesticide residues is the most widely used method of monitoring the amount of pesticide entering colonies and its change over time. In this study, we collected and analyzed pollen from 70 sites across four bee-pollinated crops over two years to characterize the variation in pesticide detection across sites, crops and at different periods during bloom. Hazard Quotient, HQ, is the most common way that pesticide residues are aggregated into a single pesticide hazard value in the current literature. Therefore, change in pesticide hazard (HQ) was quantified in composite pollen samples collected from pollen traps and in pollen color subsamples separated into pollen from the target crop being pollinated and pollen from other plant species. We used our estimates of the variation in HQ to calculate the number of sample location sites needed to detect a 5% annual change in HQ across all crops or within specific crops over a 5-year period. The number of sites required to be sampled varied by crop and year and ranged between 139 and 7194 sites, costing an estimated $129,548 and $3.35 million, respectively. The HQ values detectable for this cost would be 575 and 154. We identified additional factors that complicate the interpretation of the results as a way to evaluate changes in pest management practices at a state level. First, in all but one crop (meadowfoam), the pollen collected from outside the crop honey bee colonies were pollinating comprised a major percentage of the total pollen catch. Moreover, we found that when the overall quantity of pollen from different pollen sources was taken into account, differences in HQ among crops widened. We also found that while HQ estimates remain consistent across the bloom period for some crops, such as cherry, we observed large differences in other crops, notably meadowfoam. Overall, our results suggest the current practice of interpreting pesticides levels in pollen may come with limitations for agencies charged with improving pesticide stewardship due to the high variation associated with HQ values over time and across crops. Despite the limitations of HQ for detecting change in pesticide hazard, there remains a potential for HQ to provide feedback to regulators and scientists on field-realistic pesticide hazard within a landscape., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Carlson et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Accurate characterization of the movement of water through catchments, particularly during precipitation event response, is critical for hydrological efforts such as contaminant transport modeling or prediction of extreme flows. Abiotic hydrogeochemical tracers are commonly used to track sources and ages of surface waters but provide limited details about transit pathways or the spatial dynamics of water storage and release. Alternatively, biotic material in streams is derived from thousands of taxa originating from a variety of environments within watersheds, including groundwater, sediment, and upslope terrestrial environments, and this material can be characterized with genetic sequencing and bioinformatics. We analyzed the stable water isotopes (δ18O and δ2H) and microbiome composition (16S rRNA gene amplicon sequencing) of the Marys River of western Oregon, USA during an early season storm to describe the processes, storage, and flowpaths that shape surface water hydrology. Stable water isotopes (δ18O and δ2H) typified an event response in which stream water is composed largely of 'old' water introduced to the catchment before the storm, a common though not well understood phenomenon. In contrast, microbial biodiversity spiked during the storm, consisting of early- and late-event communities clearly distinguishable from pre-event communities. We applied concentration-discharge (cQ) analysis to individual microbial taxa and found that most Alphaproteobacteria sequences were positively correlated (i.e., were mobilized) with discharge, whereas most sequences from phyla Gammaproteobacteria and Bacteroidota were negatively correlated with discharge (i.e., were diluted). Source predictions using the prokaryote habitat preference database ProkAtlas found that freshwater-associated microbes composed a smaller fraction of the microbial community during the stream rise and a larger fraction during the recession, while soil and biofilm-associated microbes increased during the storm and remained high during recession. This suggests that the "old" water discharged during the storm was likely stored and released from, or passed through, soil- and biofilm-rich environments, demonstrating that this approach adds new, biologically derived tracer information about the hydrologic pathways active during and after this event. Overall, this study demonstrates an approach for integrating information-rich DNA into water resource investigations, incorporating tools from both hydrology and microbiology to demonstrate that microbial DNA is useful not only as an indicator of biodiversity but also functions as an innovative hydrologic tracer., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Cation conducting channelrhodopsins (ChRs) are a popular tool used in optogenetics to control the activity of excitable cells and tissues using light. ChRs with altered ion selectivity are in high demand for use in different cell types and for other specialized applications. However, a detailed mechanism of ion permeation in ChRs is not fully resolved. Here, we use complementary experimental and computational methods to uncover the mechanisms of cation transport and valence selectivity through the channelrhodopsin chimera, C1C2, in the high- and low-conducting open states. Electrophysiology measurements identified a single-residue substitution within the central gate, N297D, that increased Ca2+ permeability vs. Na+ by nearly two-fold at peak current, but less so at stationary current. We then developed molecular models of dimeric wild-type C1C2 and N297D mutant channels in both open states and calculated the PMF profiles for Na+ and Ca2+ permeation through each protein using well-tempered/multiple-walker metadynamics. Results of these studies agree well with experimental measurements and demonstrate that the pore entrance on the extracellular side differs from original predictions and is actually located in a gap between helices I and II. Cation transport occurs via a relay mechanism where cations are passed between flexible carboxylate sidechains lining the full length of the pore by sidechain swinging, like a monkey swinging on vines. In the mutant channel, residue D297 enhances Ca2+ permeability by mediating the handoff between the central and cytosolic binding sites via direct coordination and sidechain swinging. We also found that altered cation binding affinities at both the extracellular entrance and central binding sites underly the distinct transport properties of the low-conducting open state. This work significantly advances our understanding of ion selectivity and permeation in cation channelrhodopsins and provides the insights needed for successful development of new ion-selective optogenetic tools., Competing Interests: Sandia National Laboratories is a multimission laboratory managed and operated by National Technology & Engineering Solutions of Sandia, LLC, a wholly owned subsidiary of Honeywell International, Inc., for the U.S. DOE’s National Nuclear Security Administration under contract DE-NA-0003525. This does not alter our adherence to PLOS ONE policies on sharing data and materials. There are no patents, products in development or marketed products associated with this research to declare., (Copyright: © 2024 Prignano et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objective: Investigate maternal and neonatal outcomes associated with breech presentation in planned community births in the United States, including outcomes associated with types of breech presentation (i.e., frank, complete, footling/kneeling)., Design: Secondary analysis of prospective cohort data from a national perinatal data registry (MANA Stats)., Setting: Planned community birth (homes and birth centers), United States., Sample: Individuals with a term, singleton gestation (N = 71,943) planning community birth at labor onset., Methods: Descriptive statistics to calculate associations between types of breech presentation and maternal and neonatal outcomes., Main Outcome Measures: Maternal: intrapartum/postpartum transfer, hospitalization, cesarean, hemorrhage, severe perineal laceration, duration of labor stages and membrane rupture Neonatal: transfer, hospitalization, NICU admission, congenital anomalies, umbilical cord prolapse, birth injury, intrapartum/neonatal death., Results: One percent (n = 695) of individuals experienced breech birth (n = 401, 57.6% vaginally). Most fetuses presented frank breech (57%), with 19% complete, 18% footling/kneeling, and 5% unknown type of breech presentation. Among all breech labors, there were high rates of intrapartum transfer and cesarean birth compared to cephalic presentation (OR 9.0, 95% CI 7.7-10.4 and OR 18.6, 95% CI 15.9-21.7, respectively), with no substantive difference based on parity, planned site of birth, or level of care integration into the health system. For all types of breech presentations, there was increased risk for nearly all assessed neonatal outcomes including hospital transfer, NICU admission, birth injury, and umbilical cord prolapse. Breech presentation was also associated with increased risk of intrapartum/neonatal death (OR 8.5, 95% CI 4.4-16.3), even after congenital anomalies were excluded., Conclusions: All types of breech presentations in community birth settings are associated with increased risk of adverse neonatal outcomes. These research findings contribute to informed decision-making and reinforce the need for breech training and research and an increase in accessible, high-quality care for planned vaginal breech birth in US hospitals., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Schafer et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

A powerful way to predict how ecological communities will respond to future climate change is to test how they have responded to the climate of the past. We used climate oscillations including the Pacific Decadal Oscillation (PDO), North Pacific Gyre Oscillation, and El Niño Southern Oscillation (ENSO) and variation in upwelling, air temperature, and sea temperatures to test the sensitivity of nearshore rocky intertidal communities to climate variability. Prior research shows that multiple ecological processes of key taxa (growth, recruitment, and physiology) were sensitive to environmental variation during this time frame. We also investigated the effect of the concurrent sea star wasting disease outbreak in 2013-2014. We surveyed nearly 150 taxa from 11 rocky intertidal sites in Oregon and northern California annually for up to 14-years (2006-2020) to test if community structure (i.e., the abundance of functional groups) and diversity were sensitive to past environmental variation. We found little to no evidence that these communities were sensitive to annual variation in any of the environmental measures, and that each metric was associated with < 8.6% of yearly variation in community structure. Only the years elapsed since the outbreak of sea star wasting disease had a substantial effect on community structure, but in the mid-zone only where spatially dominant mussels are a main prey of the keystone predator sea star, Pisaster ochraceus. We conclude that the established sensitivity of multiple ecological processes to annual fluctuations in climate has not yet scaled up to influence community structure. Hence, the rocky intertidal system along this coastline appears resistant to the range of oceanic climate fluctuations that occurred during the study. However, given ongoing intensification of climate change and increasing frequencies of extreme events, future responses to climate change seem likely., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Gravem et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Lithified layers of complex microbial mats known as microbialites are ubiquitous in the fossil record, and modern forms are increasingly identified globally. A key challenge to developing an understanding of microbialite formation and environmental role is how to investigate complex and diverse communities in situ. We selected living, layered microbialites (stromatolites) in a peritidal environment near Schoenmakerskop, Eastern Cape, South Africa to conduct a spatial survey mapping the composition and small molecule production of the microbial communities from environmental samples. Substrate core samples were collected from nine sampling stations ranging from the upper point of the freshwater inflow to the lower marine interface where tidal overtopping takes place. Substrate cores provided material for parallel analyses of microbial community diversity by 16S rRNA gene amplicon sequencing and metabolomics using LC-MS2. Species and metabolite diversities were correlated, and prominent specialized metabolites were targeted for preliminary characterization. A new series of cyclic hexadepsipeptides, named ibhayipeptolides, was most abundant in substrate cores of submerged microbialites. These results demonstrate the detection and identification of metabolites from mass-limited environmental samples and contribute knowledge about microbialite chemistry and biology, which facilitates future targeted studies of specialized metabolite function and biosynthesis., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Neuhaus et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

This study presents the complete genome sequence of a novel nege-like virus identified in whiteflies (Bemisia tabaci MEAM1), provisionally designated as whitefly negevirus 1 (WfNgV1). The virus possesses a single-stranded RNA genome comprising 11,848 nucleotides, organized into four open reading frames (ORFs). These ORFs encode the putative RNA-dependent-RNA-polymerase (RdRp, ORF 1), a glycoprotein (ORF 2), a structural protein with homology to those in the SP24 family, (ORF 3), and a protein of unknown function (ORF 4). Phylogenetic analysis focusing on RdRp and SP24 amino acid sequences revealed a close relationship between WfNgV1 and Bemisia tabaci negevirus 1, a negevirus sequence recently discovered in whiteflies from Israel. Both viruses form a clade sharing a most recent common ancestor with the proposed nelorpivirus and centivirus taxa. The putative glycoprotein from ORF 2 and SP24 (ORF 3) of WfNgV1 exhibit the characteristic topologies previously reported for negevirus counterparts. This marks the first reported negevirus-like sequence from whiteflies in the Americas., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Quito-Avila et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The impressive †Oncorhynchus rastrosus of the Pacific Northwest's Miocene and Pliocene eras was the largest salmonid ever to live. It sported a hypertrophied premaxilla with a pair of enlarged teeth which the original describers reconstructed as projecting ventrally into the mouth, leading them to assign the species to "Smilodonichthys," a genus now in synonymy. Through CT reconstruction of the holotype and newly collected specimens, we demonstrate that the famed teeth projected laterally like tusks, not ventrally like sabers or fangs. We also expand the original description to characterize sexual dimorphism in mature, breeding individuals. Male and female †Oncorhynchus rastrosus differ in the form of the vomer, rostro-dermethmoid-supraethmoid, and dentary, much as do other extant species of Oncorhynchus. Male specimens possess a more elongate vomer than do females, and female vomers have concave ventral surfaces and prominent median dorsal keels. The dentary of females has no evidence of a kype, though some specimens of †O. rastrosus have a non-uniform density mesial to the tooth bed, which we interpret as a male kype. Unlike extant Oncorhynchus, male and female †O. rastrosus do not differ in premaxilla shape. Because male and females possess hypertrophied premaxillae and lateral premaxillary spikes, the former common name "Sabertoothed Salmon" no longer reflects our understanding of the species' morphology. Accordingly, we redub †O. rastrosus the Spike-Toothed Salmon and postulate that its spikes were multifunctional, serving as defense against predators, in agonism against conspecifics, and as a practical aid to nest construction., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Claeson et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Cooperation via shared public goods is ubiquitous in nature, however, noncontributing social cheaters can exploit the public goods provided by cooperating individuals to gain a fitness advantage. Theory predicts that this dynamic can cause a Tragedy of the Commons, and in particular, a 'Collapsing' Tragedy defined as the extinction of the entire population if the public good is essential. However, there is little empirical evidence of the Collapsing Tragedy in evolutionary biology. Here, we experimentally demonstrate this outcome in a microbial model system, the public good-producing bacterium Pseudomonas aeruginosa grown in a continuous-culture chemostat. In a growth medium that requires extracellular protein digestion, we find that P. aeruginosa populations maintain a high density when entirely composed of cooperating, protease-producing cells but completely collapse when non-producing cheater cells are introduced. We formulate a mechanistic mathematical model that recapitulates experimental observations and suggests key parameters, such as the dilution rate and the cost of public good production, that define the stability of cooperative behavior. We combine model prediction with experimental validation to explain striking differences in the long-term cheater trajectories of replicate cocultures through mutational events that increase cheater fitness. Taken together, our integrated empirical and theoretical approach validates and parametrizes the Collapsing Tragedy in a microbial population, and provides a quantitative, mechanistic framework for generating testable predictions of social behavior., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Lynn et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Beginning in December 2018, increased numbers of gray whale (Eschrichtius robustus) strandings were reported along the west coast of Mexico, the United States, and Canada, prompting declaration of a gray whale Unusual Mortality Event (UME) by the United States National Marine Fisheries Service. Although strandings declined in 2020 and 2021 from a peak in 2019, the UME is still ongoing as of fall 2023. Between 17 December 2018 and 31 December 2021, 503 animals stranded along the west coast of North America, with 226 strandings in Mexico, 71 in California, 12 in Oregon, 56 in Washington, 21 in British Columbia, and 117 in Alaska. These included 187 males, 167 females, and 149 whales of undetermined sex; and 193 adults, 194 subadults, 40 calves, 1 fetus, and 75 whales of undetermined age class. We report on 61 of the 503 carcasses (12%) that had external and internal gross necropsy and/or histopathology data: of these 61 whales, findings that contributed to death were identified in 33 (54%) whales. Sixteen of the 61 (26%) were severely emaciated. Gross lesions of blunt force trauma consistent with vessel strike were identified in 11 of the 61 animals (18%), only two of which were emaciated. Two whales (3%) were entangled at time of death, and one died from entrapment. Signs of killer whale (Orcinus orca) interaction were documented in 19 of the 61 animals; five were deemed from recent interactions and three (5%) likely contributed to mortality. A specific cause of death could not be identified in 28 of 61 whales (46%). Additionally, logistical challenges and the advanced state of decomposition of most examined carcasses precluded detection of potential infectious or toxic causes of morbidity or mortality. Up to 2016, the eastern North Pacific population of gray whale population had generally been increasing since the cessation of historic whaling and a prior UME in 1999-2000. However, recent abundance and calf production estimates have declined, a trend that overlaps the current UME. The relative contributions of carrying capacity, environmental change, prey shifts, and infectious, toxic, and other processes to the increased gray whale mortalities have not yet been resolved. Nevertheless, the marked temporal increase in strandings, including findings of malnutrition in some of the whales, along with low calf production, likely represent consequences of complex and dynamic ecological interactions in the ocean impacting the population., Competing Interests: The authors have no competing interests., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Inundative biological control (biocontrol) efforts in pest management lead to the mass distribution of commercialized biocontrol agents. Many 'biocontrol gone awry' incidents have resulted in disastrous biodiversity impacts, leading to increased scrutiny of biocontrol efforts. The nematode Phasmarhabditis hermaphrodita is sold as a biocontrol agent on three continents and targets pest gastropods such as Deroceras reticulatum, the Grey Field Slug; P. hermaphrodita is not presently approved for use in the United States. Investigations into the potential for P. hermaphrodita to infect non-target gastropod species of conservation relevance, however, are limited. We examined the effects of three strains of P. hermaphrodita on mortality in Monadenia fidelis, the Pacific Sideband, a snail species endemic to the Pacific Northwest of North America, in laboratory conditions. Across a 71-day laboratory infectivity assay, snails exposed to each of the three nematode strains, each analyzed at two doses, experienced a mean 50% mortality by days 20-42. All nematode-treated snails were dead by the end of the study. By contrast, 30/30 water-control snails experienced no mortality. Nematodes killed smaller, juvenile-stage snails significantly faster than those in larger and more developmentally advanced hosts. Our results provide direct evidence that the biocontrol nematode P. hermaphrodita infects and kills M. fidelis, a non-target gastropod species endemic to the Pacific Northwest, in laboratory conditions. This study suggests that introduction of P. hermaphrodita to new ecosystems might negatively impact endemic gastropod biodiversity and advocates for further investigation of non-target effects, including in conditions closer to the natural environments of non-target species., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Denver et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Syntax, the combination of meaning-devoid phonemes into meaningful words, which in turn are combined in structurally and semantically complex sentences, is fundamental to the unlimited expressiveness of human languages. Studying the functions of call combinations in non-human species provides insights into the evolution of such syntactic capabilities. Here, we investigated the combination of high amplitude broadband calls with low frequency rumble vocalizations in a highly social species, the African forest elephant Loxodonta cyclotis. Rumbles play an integral role in coordinating social interactions by transmitting socially relevant information, including individual identity. By contrast, broadband calls, such as roars, are thought to function as signals of distress and urgency as they are typically produced in situations of high emotional intensity. Functional changes associated with the combination of these calls remain little understood. We found that call combinations were produced by all age-sex classes but were most prevalent in immature individuals. We found that rumbles used singularly occurred in all five investigated social contexts, whereas single broadband calls were restricted to two resource-related contexts. Call combinations also occurred in all five contexts, suggesting an increase in the functional use of broadband calls when combined with rumbles, analogous to the generativity brought about through syntax in human speech. Moreover, combining calls appeared to lead to functional shifts towards high-stake contexts. Call combinations were more likely in competition contexts compared to single rumbles, and more likely in separation contexts compared to single broadband calls. We suggest that call combination in forest elephants may aide to reduce message ambiguity in high-stake situation by simultaneously communicating distress and individual identity, which may be critical to secure access to resources, reduce the risk of injury and to reunite with or recruit the support of the family group., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Hedwig, Kohlberg. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Life tables are one of the most common tools to describe the biology of insect species and their response to environmental conditions. Although the benefits of life tables are beyond question, we raise some doubts about the completeness of the information reported in life tables. To substantiate these doubts, we consider a case study (Corcyra cephalonica) for which the raw dataset is available. The data suggest that the Gaussian approximation of the development times which is implied by the average and standard error usually reported in life tables does not describe reliably the actual distribution of the data which can be misleading and hide interesting biological aspects. Furthermore, it can be risky when life table data are used to build models to predict the demographic changes of the population. The present study highlights this aspect by comparing the impulse response generated by the raw data and by its Gaussian approximation based on the mean and the standard error. The conclusions of this paper highlight: i) the importance of adding more information to life tables and, ii) the role of raw data to ensure the completeness of this kind of studies. Given the importance of raw data, we also point out the need for further developments of a standard in the community for sharing and analysing data of life tables experiments., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Rossini et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

It was recently reported that values of the transition heat capacities, as measured by differential scanning calorimetry, for two globular proteins and a short DNA hairpin in NaCl buffer are essentially equivalent, at equal concentrations (mg/mL). To validate the broad applicability of this phenomenon, additional evidence for this equivalence is presented that reveals it does not depend on DNA sequence, buffer salt, or transition temperature (Tm). Based on the equivalence of transition heat capacities, a calorimetric method was devised to determine protein concentrations in pure and complex solutions. The scheme uses direct comparisons between the thermodynamic stability of a short DNA hairpin standard of known concentration, and thermodynamic stability of protein solutions of unknown concentrations. Sequences of two DNA hairpins were designed to confer a near 20°C difference in their Tm values. In all cases, evaluated protein concentrations determined from the DNA standard curves agreed with the UV-Vis concentration for monomeric proteins. For multimeric proteins evaluated concentrations were greater than determined by UV-Vis suggesting the calorimetric approach can also be an indicator of molecular stoichiometry., Competing Interests: We have read the journal’s policy and the authors of this manuscript have the following competing interests: M.W.E. and A.S.B. are co‐inventors on several patent applications assigned to Portland State University. M.W.E. and A.S.B. are also founders of a biotechnology company, ThermoCap Laboratories Inc, that is involved in development of these technologies. This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2024 Eskew et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

This study presents the status and trends of long-term monitoring of the elemental concentrations of zinc (Zn), lead (Pb), and cadmium (Cd) in Hylocomium splendens moss tissue in Cape Krusenstern National Monument (CAKR), Alaska, adjacent to the Red Dog Mine haul road. Spatial patterns of the deposition of these metals were re-assessed for the period from 2006-2017 following an identical study that assessed trends between 2001-2006. In contrast to the widespread and steep declines in Zn and Pb levels throughout most of the study area between 2001-2006, this study showed more mixed results for 2006-2017. At distances within 100 m of the haul road, only Pb decreased between 2006-2017. At distances between 100-5,000 m, however, both Zn and Cd decreased between 2006-2017, with high probabilities of decrease and percent decreases of 11-20% and 46-52% respectively. Lead did not decrease in any of the more distant areas. Following earlier work on lichen species richness in the study area, it appears that 2017 Zn levels are approaching those associated with "background" lichen species richness throughout a relatively large proportion of the study area at least 2,000 m from the haul road and several km from the port site. The findings in this study may be used to plan additional mitigation measures to reduce Zn deposition related to impacts on lichen communities., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

The aims of this study were to (1) establish the intrapopulation and seasonal variation of Juniperus excelsа essential oil (EO); (2) compare the J. excelsa concrete and resinoid composition with its EO composition; and (3) investigate the potential herbicidal activity of J. excelsa EO against seeds of Papaver rhoeas L., Consolida orientalis (J.Gay) Schrödinger, Anthemis arvensis L., Avena fatua L., and Agrostemma githago L. Four independent studies were performed to meet these objectives. Twenty-eight individual trees were analyzed from two populations to establish intrapopulation and interpopulation variability of EOs yield and composition. In the seasonal dynamic study of leaf EO, samples from the same three trees and in the same population were collected in January, March, May, July, October, and December and their EO yield and composition determined. The EOs (intrapopulation and seasonal) were extracted by hydrodistillation, while the EO for the herbicidal test was obtained by steam distillation in a semi-commercial (SCom) apparatus. Overall, the EO yield varied significantly from 0.93% to 2.57%. α-Pinene (8.85-35.94%), limonene (11.81-50.08%), and cedrol (3.41-34.29%) were the predominant EO compounds in all samples (intrapopulation variability); however, trans-2,4-decadienol and β-caryophyllene were predominant in some individual trees. Four chemical groups were identified in the samples collected from two natural populations (intrapopulation). This is the first report on the compositions of J. excelsa concrete and resinoid. Cedrol (15.39%), 7-hydroxy-4-methyl-coumarin (17.63%), 1-octacosanol (36.85%), tritriacontane (16.08%), and tiacontanoic acid were the main compounds in the concrete and resinoid. Juniperus excelsa EO suppressed seed germination and seedling growth of P. rhoeas, C. orientalis, A. arvensis, A. fatua, and A. githago, demonstrating its potential to be used for the development of new biopesticides. The highest EO yield with high content of limonene and cedrol was obtained from samples harvested during the winter months (December, January, and March)., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Semerdjieva et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The Northern California Current is a highly productive marine upwelling ecosystem that is economically and ecologically important. It is home to both commercially harvested species and those that are federally listed under the U.S. Endangered Species Act. Recently, there has been a global shift from single-species fisheries management to ecosystem-based fisheries management, which acknowledges that more complex dynamics can reverberate through a food web. Here, we have integrated new research into an end-to-end ecosystem model (i.e., physics to fisheries) using data from long-term ocean surveys, phytoplankton satellite imagery paired with a vertically generalized production model, a recently assembled diet database, fishery catch information, species distribution models, and existing literature. This spatially-explicit model includes 90 living and detrital functional groups ranging from phytoplankton, krill, and forage fish to salmon, seabirds, and marine mammals, and nine fisheries that occur off the coast of Washington, Oregon, and Northern California. This model was updated from previous regional models to account for more recent changes in the Northern California Current (e.g., increases in market squid and some gelatinous zooplankton such as pyrosomes and salps), to expand the previous domain to increase the spatial resolution, to include data from previously unincorporated surveys, and to add improved characterization of endangered species, such as Chinook salmon (Oncorhynchus tshawytscha) and southern resident killer whales (Orcinus orca). Our model is mass-balanced, ecologically plausible, without extinctions, and stable over 150-year simulations. Ammonium and nitrate availability, total primary production rates, and model-derived phytoplankton time series are within realistic ranges. As we move towards holistic ecosystem-based fisheries management, we must continue to openly and collaboratively integrate our disparate datasets and collective knowledge to solve the intricate problems we face. As a tool for future research, we provide the data and code to use our ecosystem model., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Sagebrush steppe is one of the most threatened ecosystems in North America. Adult density of songbirds within sagebrush steppe is a metric used to evaluate conservation actions. However, relying on only adult density to guide conservation may be misleading. Information on how conservation actions influence the nest density and nest survival of songbird species, in addition to adult density, are needed. We evaluated the relationships between nest density, nest survival, and adult density of Brewer's sparrow (Spizella breweri) and vesper sparrow (Pooecetes gramineus) over 3 breeding seasons in central Montana. Our findings suggest that adult pairs of both species were often present in higher numbers than nests, and this relationship was most prominent for Brewer's sparrows. However, our results do not support density dependence when considering nest survival. This discrepancy suggests that songbirds may not breed every year and that density dependence may be operating on nest densities within these populations differently than we examined. This study provides information on relationships between population demographics for 2 songbird species in grazed sagebrush steppe that will improve monitoring and management activities of conservation efforts., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Ruth et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Yeast-derived products have become more of an interest in the poultry industry as of late because of their use in modulating the gastrointestinal tract (GIT) microbiome to both improve production parameters and prevent infection. This study aimed to evaluate the effects of various yeast-derived products on Salmonella enterica inoculation in un in vitro rooster cecal incubations and associated effects on the cecal microbiome. Cecal contents were obtained from 53-wk old White Leghorn H & N Nick Chick roosters (n = 3) fed a wheat-based, commercial-type basal diet. Cecal contents were diluted 1:3000 in anaerobic dilution solution (ADS) in an anaerobic chamber, with 20 mL aliquoted to each serum bottle. There were three controls (n = 3): basal diet only, diluted cecal contents only, and basal diet and diluted cecal contents; and five treatments containing the basal diet and diluted cecal contents (n = 3): Citristim® (ADM), ImmunoWall® (ICC), Maxi-Gen Plus® (CBS Bio Platforms), Hilyses® (ICC), and Original XPC® (Diamond V). All treatments were applied at a rate of 2.5 kg/tonne or less. All groups were inoculated with a nalidixic acid-resistant strain of Salmonella Enteritidis at 10^7 CFU/mL and incubated at 37 deg C. Samples were collected at 0, 24, and 48 h for S. Enteritidis enumeration and 16S rDNA microbial sequencing. Salmonella data were log-transformed and analyzed in a two-way ANOVA with means separated using Tukey's HSD (P≤0.05). Genomic DNA was extracted, and resulting libraries were prepared and sequenced using an Illumina MiSeq. Sequencing data were analyzed in QIIME2 (2021.4) with diversity metrics (alpha and beta), and an analysis of the composition of microbiomes (ANCOM) was performed. Main effects were considered significant at P≤0.05, with pairwise differences considered significant at Q≤0.05. There was an interaction of treatment and time on the enumeration of Salmonella where treatments of Citristim, Immunowall, Hilyses, and XPC reduced Salmonella by 1 log CFU/mL compared to the controls. At 48 h, each yeast product treatment reduced Salmonella by 3 log CFU/mL compared to the controls. There was no main effect of treatment on the alpha diversity metrics, richness, or evenness (P > 0.05). Treatment affected the beta diversity, abundance, and phylogenetic differences, but there were no pairwise differences (P>0.05, Q>0.05). Using ANCOM at the genus level, the taxa Synergistes, Alloprevotella, Sutterella, and Megasphaera abundance were significantly different (W = 154,147,145,140, respectively). These results demonstrate the potential of these yeast-derived products to reduce foodborne pathogens, such as Salmonella Enteriditis, in vitro, without negatively disrupting the cecal microbiome., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Costello et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Methods and models to design, prioritize and evaluate fuel break networks have potential application in many fire-prone ecosystems where major increases in fuel management investments are planned in response to growing incidence of wildfires. A key question facing managers is how to scale treatments into manageable project areas that meet operational and administrative constraints, and then prioritize their implementation over time to maximize fire management outcomes. We developed and tested a spatial modeling system to optimize the implementation of a proposed 3,538 km fuel break network and explore tradeoffs between two implementation strategies on a 0.5 million ha national forest in the western US. We segmented the network into 2,766 treatment units and used a spatial optimization model to compare linear versus radial project implementation geometries. We hypothesized that linear projects were more efficient at intercepting individual fire events over larger spatial domains, whereas radial projects conferred a higher level of network redundancy in terms of the length of the fuel break exposed to fires. We simulated implementation of the alternative project geometries and then examined fuel break-wildfire spatial interactions using a library of simulated fires developed in prior work. The results supported the hypothesis, with linear projects exhibiting substantially greater efficiency in terms of intercepting fires over larger areas, whereas radial projects had a higher interception length given a fire encountered a project. Adding economic objectives made it more difficult to obtain alternative project geometries, but substantially increased net revenue from harvested trees. We discuss how the model and results can be used to further understand decision tradeoffs and optimize the implementation of planned fuel break networks in conjunction with landscape conservation, protection, and restoration management in fire prone regions., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Over 2 million people in North America suffer from inflammatory bowel disease (IBD), a chronic and idiopathic inflammatory condition. While previous research has primarily focused on studying immune cells as a cause and therapeutic target for IBD, recent findings suggest that non-immune cells may also play a crucial role in mediating cytokine and chemokine signaling, and therefore IBD symptoms. In this study, we developed an organ-on-a-chip co-culture model of Caco2 epithelial and HUVEC endothelial cells and induced inflammation using pro-inflammatory cytokines TNF-α and IFN-γ. We tested different concentration ranges and delivery orientations (apical vs. basal) to develop a consistently inducible inflammatory response model. We then measured pro-inflammatory cytokines and chemokines IL-6, IL-8, and CXCL-10, as well as epithelial barrier integrity. Our results indicate that this model 1. induces IBD-like cytokine secretion in non-immune cells and 2. decreases barrier integrity, making it a feasible and reliable model to test the direct actions of potential anti-inflammatory therapeutics on epithelial and endothelial cells., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: MMD has financial interests in NeuroBiome LLC, Microbiome Engineering LLC and Second genome Inc. NeuroBiome LLC develops technology related to this research. The conduct, outcomes, or reporting of this research could benefit NeuroBiome LLC and could potentially benefit MMD. This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2023 Tataru et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Introduction: Centranthus kellereri is a Bulgarian endemic plant species, found only in two locations in the world: The Balkans Mountains (Stara Planina), above the town of Vratsa, and The Pirin Mountains, above the town of Bansko, Bulgaria. Being endemic and endangered species precluded any significant research on it. The hypothesis of this study was that the populations of C. kellereri may represent genetically, phytochemically, and morphologically distinct forms and these will differentiate from C. ruber. Furthermore, C. kellereri possibly imperfect embryology may preclude its more widespread distribution under natural conditions., Results: This study revealed the phytochemical profile, antioxidant activity, embryology, surface microstructural morphological traits, and genetic differences between the C. kellereri plants from the only two natural populations and compares them to the ones of the related and better-known plant C. ruber. The essential oil (EO) content in aboveground plant parts and in roots was generally low and the EO composition varied significantly as a function of plant part, year of sampling, location, and species. Methylvaleric acid was a major EO constituent in the C. kellereri EO, ranging between 60.2% and 71.7% of the total EO. The EO included monoterpenes, sequiterpenes, long-chain alkanes and fatty acids. Phytochemical analyses of plant tissue revealed the occurrence of 32 compounds that were tentatively identified as 6 simple phenolics, 18 flavonoids, 1 quinone, 1 lipid, 1 alkaloid, 2 diterpenes, and 3 triterpenes. There were differences in detected compounds between the C. kellereri plants at the two locations and between the roots and shoots in both species. The total phenols and flavonoids varied between the two species but were also dissimilar between the plants from the two populations of C. kelleri. Free radical scavenging activity, measured with ABTS and DPPH in aqueous and methanol extracts, had similar values; however, overall, C. kellereri from Vratsa showed the highest antioxidant activity while C. ruber had the lowest activity. Genetic analyses showed a clear differentiation between C. kellereri and C. ruber, and between the two populations of C. kellereri. Embryological studies revealed the peculiarities of the male and female generative spheres of the two species that were defined as being sexually reproducing. The pollen had high viability; however, the low viability of seeds demonstrated possible high sensitivity of C. kellereri to the environmental conditions, perhaps the main factor modifying and restricting the population sizes. The SEM analyses exposed differences in surface microstructural traits between the species (C. kellereri and C. ruber) but also between the two populations of C. kellereri. The observed dissimilarities in genetic makeup, micromorphological characteristics, and phytochemical composition strongly indicate that the two populations can be classified as distinct subspecies or varieties of C. kellereri; var. pirinensis and var. balkanensis. Further research is needed to introduce C. kellereri into culture and develop it as a high-value specialty crop or ornamental in order to conserve C. kellereri natural populations. C. kellereri may be utilized as a source for phytochemicals of interest and as an ornamental plant like C. ruber; however, it may have a greater environmental plasticity and adaptation as evidenced by its current locations., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Zheljazkov et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Nonalcoholic fatty liver disease (NAFLD) is a global health problem. Identification of factors contributing to the onset and progression of NAFLD have the potential to direct novel strategies to combat NAFLD., Methods: We examined the time course of western diet (WD)-induced NAFLD and its progression to nonalcoholic steatohepatitis (NASH) in age-matched female and male Ldlr-/- mice, with time-points at 1, 4, 8, 20 and 40 weeks on the WD. Controls included Ldlr-/- mice maintained on a purified low-fat diet (LFD) for 1 and 40 weeks. The approach included quantitation of anthropometric, plasma and liver markers of disease, plus hepatic histology, lipids, oxylipins, gene expression and selected metabolites., Results: One week of feeding the WD caused a significant reduction in hepatic essential fatty acids (EFAs: 18:2, ω6, 18:3, ω3) which preceded the decline in many C20-22 ω3 and ω6 polyunsaturated fatty acids (PUFA) and PUFA-derived oxylipins after 4 weeks on the WD. In addition, expression of hepatic inflammation markers (CD40, CD44, Mcp1, Nlrp3, TLR2, TLR4, Trem2) increased significantly in both female & male mice after one week on the WD. These markers continued to increase over the 40-week WD feeding study. WD effects on hepatic EFA and inflammation preceded all significant WD-induced changes in body weight, insulin resistance (HOMA-IR), oxidative stress status (GSH/GSSG ratio) and histological and gene expression markers of macrosteatosis, extracellular matrix remodeling and fibrosis., Conclusions: Our findings establish that feeding Ldlr-/- mice the WD rapidly lowered hepatic EFAs and induced key inflammatory markers linked to NASH. Since EFAs have an established role in inflammation and hepatic inflammation plays a major role in NASH, we suggest that early clinical assessment of EFA status and correcting EFA deficiencies may be useful in reducing NASH severity., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Spooner et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Nutritional programming is the idea that early nutrient contributions can influence organismal structure or function and is documented in a variety of vertebrates, yet studies in fish are largely lacking. Tilapia are an important foodfish, with global production having increased rapidly since the 1990s. They exhibit high disease-resistance and grow well on formulated feeds which makes them an ideal aquaculture species, however incorporating high quality proteins into feeds can be costly. As feed constitutes 50-70% of total production costs in aquaculture, reducing protein content could curb these costs and increase revenue. Thus, we examined the effects of feeding Nile tilapia (O. niloticus) fry a restricted protein diet for the first 7-21 days on growth, gut microbial flora, and the intestinal transcriptome. Fish were fed either a 25% restricted or 48% control crude protein starter (ST) diet for up to 21 days and then switched to a 25% or 38% control crude protein growout (GO) diet. Fish fed a 25% ST diet for 14 days followed by a 38% GO diet had significantly higher lengths and weights and better feed efficiency than fish fed the control 48% ST and 38% GO diet after 56 days of culture. Growth of fry on the 25% ST, 7-day/38% GO and the 25% ST,7-day/25% GO diets did not differ from the those fed the control protein diets, while fish fed the 25% ST diet for 21 days had significantly lower growth and survival rates. We observed no significant differences in either alpha or beta diversity of the gut microbial flora between diets, however species richness (Shannon Index) was higher in fry fed the 25% protein ST diet regardless of the GO diet. Similarly, fish fed the 25% ST diet for 14 days followed by the 38% GO diet had minimal changes to the intestinal transcriptome relative to fish fed the control 48% ST and 38% GO diet. However, those fed 25% ST and GO diets for the entire 56 days exhibited substantial differences in the gut transcriptome from other groups showing gene expression profiles characteristic of detrimental changes to gut physiology, protein metabolism and immune function. Results suggest protein restriction for up to 14 days early in development leads to enhanced growth and feed efficiency with minimal effects on gut microbes or intestinal function. Protein restriction beyond this period appears detrimental to fish growth and health as underscored by expression of disease related genes and higher mortality rates., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Deck et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Despite established relationships between diabetic status and an increased risk for COVID-19 severe outcomes, there is a limited number of studies examining the relationships between diabetes complications and COVID-19-related risks. We use the Adapted Diabetes Complications Severity Index to define seven diabetes complications. We aim to understand the risk for COVID-19 infection, hospitalization, mortality, and longer length of stay of diabetes patients with complications., Methods: We perform a retrospective case-control study using Electronic Health Records (EHRs) to measure differences in the risks for COVID-19 severe outcomes amongst those with diabetes complications. Using multiple logistic regression, we calculate adjusted odds ratios (OR) for COVID-19 infection, hospitalization, and in-hospital mortality of the case group (patients with diabetes complications) compared to a control group (patients without diabetes). We also calculate adjusted mean difference in length of stay between the case and control groups using multiple linear regression., Results: Adjusting demographics and comorbidities, diabetes patients with renal complications have the highest odds for COVID-19 infection (OR = 1.85, 95% CI = [1.71, 1.99]) while those with metabolic complications have the highest odds for COVID-19 hospitalization (OR = 5.58, 95% CI = [3.54, 8.77]) and in-hospital mortality (OR = 2.41, 95% CI = [1.35, 4.31]). The adjusted mean difference (MD) of hospital length-of-stay for diabetes patients, especially those with cardiovascular (MD = 0.94, 95% CI = [0.17, 1.71]) or peripheral vascular (MD = 1.72, 95% CI = [0.84, 2.60]) complications, is significantly higher than non-diabetes patients. African American patients have higher odds for COVID-19 infection (OR = 1.79, 95% CI = [1.66, 1.92]) and hospitalization (OR = 1.62, 95% CI = [1.39, 1.90]) than White patients in the general diabetes population. However, White diabetes patients have higher odds for COVID-19 in-hospital mortality. Hispanic patients have higher odds for COVID-19 infection (OR = 2.86, 95% CI = [2.42, 3.38]) and shorter mean length of hospital stay than non-Hispanic patients in the general diabetes population. Although there is no significant difference in the odds for COVID-19 hospitalization and in-hospital mortality between Hispanic and non-Hispanic patients in the general diabetes population, Hispanic patients have higher odds for COVID-19 hospitalization (OR = 1.83, 95% CI = [1.16, 2.89]) and in-hospital mortality (OR = 3.69, 95% CI = [1.18, 11.50]) in the diabetes population with no complications., Conclusions: The presence of diabetes complications increases the risks of COVID-19 infection, hospitalization, and worse health outcomes with respect to in-hospital mortality and longer hospital length of stay. We show the presence of health disparities in COVID-19 outcomes across demographic groups in our diabetes population. One such disparity is that African American and Hispanic diabetes patients have higher odds of COVID-19 infection than White and Non-Hispanic diabetes patients, respectively. Furthermore, Hispanic patients might have less access to the hospital care compared to non-Hispanic patients when longer hospitalizations are needed due to their diabetes complications. Finally, diabetes complications, which are generally associated with worse COVID-19 outcomes, might be predominantly determining the COVID-19 severity in those infected patients resulting in less demographic differences in COVID-19 hospitalization and in-hospital mortality., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Paramita et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The plasticity of some coral-associated microbial communities under stressors like warming and ocean acidification suggests the microbiome has a role in the acclimatization of corals to future ocean conditions. Here, we evaluated the acclimatization potential of coral-associated microbial communities of four Hawaiian coral species (Porites compressa, Porites lobata, Montipora capitata, and Pocillopora acuta) over 22-month mesocosm experiment. The corals were exposed to one of four treatments: control, ocean acidification, ocean warming, or combined future ocean conditions. Over the 22-month study, 33-67% of corals died or experienced a loss of most live tissue coverage in the ocean warming and future ocean treatments while only 0-10% died in the ocean acidification and control. Among the survivors, coral-associated microbial communities responded to the chronic future ocean treatment in one of two ways: (1) microbial communities differed between the control and future ocean treatment, suggesting the potential capacity for acclimatization, or (2) microbial communities did not significantly differ between the control and future ocean treatment. The first strategy was observed in both Porites species and was associated with higher survivorship compared to M. capitata and P. acuta which exhibited the second strategy. Interestingly, the microbial community responses to chronic stressors were independent of coral physiology. These findings indicate acclimatization of microbial communities may confer resilience in some species of corals to chronic warming associated with climate change. However, M. capitata genets that survived the future ocean treatment hosted significantly different microbial communities from those that died, suggesting the microbial communities of the survivors conferred some resilience. Thus, even among coral species with inflexible microbial communities, some individuals may already be tolerant to future ocean conditions. These findings suggest that coral-associated microbial communities could play an important role in the persistence of some corals and underlie climate change-driven shifts in coral community composition., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Price et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Detection and identification of species, subspecies or stocks of whales, dolphins and porpoises at sea remain challenging, particularly for cryptic or elusive species like beaked whales (Family: Ziphiidae). Here we investigated the potential for using an acoustically assisted sampling design to collect environmental (e)DNA from beaked whales on the U.S. Navy's Atlantic Undersea Test and Evaluation Center (AUTEC) in The Bahamas. During 12 days of August 2019, we conducted 9 small-boat surveys and collected 56 samples of seawater (paired subsamples of 1L each, including controls) using both a spatial collection design in the absence of visual confirmation of whales, and a serial collection design in the proximity of whales at the surface. There were 7 sightings of whales, including 11 Blainville's beaked whales (Mesoplodon densirostris). All whales were located initially with the assistance of information from a bottom-mounted acoustic array available on the AUTEC range. Quantification by droplet digital (dd)PCR from the four spatial design collections showed no samples of eDNA above the threshold of detection and none of these 20 samples yielded amplicons for conventional or next-generation sequencing. Quantification of the 31 samples from four serial collections identified 11 likely positive detections. eDNA barcoding by conventional sequencing and eDNA metabarcoding by next-generation sequencing confirmed species identification for 9 samples from three of the four serial collections. We further resolved five intra-specific variants (i.e., haplotypes), two of which showed an exact match to previously published haplotypes and three that have not been reported previously to the international repository, GenBank. A minimum spanning network of the five eDNA haplotypes, with all other published haplotypes of Blainville's beaked whales, suggested the potential for further resolution of differences between oceanic populations., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Baker et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The decline in salmon and steelhead populations in the Columbia River Basin has been well documented, as have the decades-long, $9 billion restoration spending efforts by federal and state agencies. These efforts are mainly tied to Endangered Species Act (ESA) mandates for recovery of wild, naturally-spawning threatened or endangered fish species. The impact of these efforts remains poorly understood; many observers, including the federal courts, have long been concerned by the lack of evidence of recovery. Most studies evaluating restoration efforts have examined individual projects for specific species, reaches, or life stages, which limits the ability to make broad inferences at the basin level. There is a need to ask: is there evidence of an overall increase in wild fish abundance associated with the totality of these recovery efforts? To that end, the current study estimates fixed-effects panel regression models of adult returns of four species. Results indicate that restoration spending combined with hatchery production are associated with substantial increases in returning adult fish. Evidence of benefits to wild fish alone, however, require indirect approaches given the commingling of restoration spending with spending on hatchery releases, the impacts of spending on hatchery fish survival, and the density dependence effects of hatchery releases. To accomplish this, the models' predicted adult returns (both hatchery and wild fish) attributed to both spending and hatchery releases are compared to independent estimates of returning hatchery fish based on hatchery survival estimates (smolt-to-adult ratios). The comparison finds the model-predicted levels of adult returns due to spending and hatchery releases do not exceed the survival-rate based estimates for hatcheries alone, so that we are unable to reject the hypothesis of no benefits to wild fish from the restoration spending., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

Objective: This study aimed to assess changes in social contact with family, friends and healthcare providers, as well as social participation in working, volunteering, religious services and other organized activities, among older adults during the COVID-19 pandemic while examining the role of pre-COVID sociodemographic characteristics or cognitive and physical limitations in changes in social contact and participation., Methods: We conducted secondary data analyses in the National Health and Aging Trends Study (NHATS) COVID-19 questionnaire, collected in 2020 during a period of workplace closures and social distancing guidelines. We linked data to pre-COVID sociodemographic and medical information collected in 2019 before COVID interrupted social life. The frequency of participants' social contact and social participation prior to and during the COVID-19 pandemic, were compared using paired t-tests for summed scores. Multivariate linear regression was used to relate participants' socio-demographic, prior physical and cognitive performance with levels of social contact and participation and with changes reported during the COVID-19 pandemic., Results: In total, results from 2,486 eligible participants revealed that COVID-19 was associated with decreased social contact among family and friends (change: -0.62; SE: 0.06; p<0.0001), and social participation among older adults (change: -0.58; SE: 0.02; p<0.0001). Pre-COVID characteristics including older age, lower educational attainment, poorer physical performance, and more chronic conditions were associated with lower social contact and social participation and with how older adults adapted their social lives during the COVID-19 pandemic., Discussion: These results emphasize the importance of increasing digital inclusion for older adults in a major crisis., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Zhang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Sea star wasting syndrome (SSWS) can cause widespread mortality in starfish populations as well as long-lasting changes to benthic community structure and dynamics. SSWS symptoms have been documented in numerous species and locations around the world, but to date there is only one record of SSWS from the Antarctic and this outbreak was associated with volcanically-driven high temperature anomalies. Here we report outbreaks of SSWS-like symptoms that affected ~30% of individuals of Odontaster validus at two different sites in McMurdo Sound, Antarctica in 2019 and 2022. Unlike many SSWS events in other parts of the world, these outbreaks were not associated with anomalously warm temperatures. Instead, we suggest they may have been triggered by high nutrient input events on a local scale. Although the exact cause of these outbreaks is not known, these findings are of great concern because of the keystone role of O. validus and the slow recovery rate of Antarctic benthic ecosystems to environmental stressors., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Moran et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Tobacco smoking imposes a staggering burden on public health, underscoring the urgency of developing a deeper understanding of the processes that maintain addiction. Clinical and experience-sampling data highlight the importance of anxious withdrawal symptoms, but the underlying neurobiology has remained elusive. Mechanistic work in animals implicates the central extended amygdala (EAc)-including the central nucleus of the amygdala and the neighboring bed nucleus of the stria terminalis-but the translational relevance of these discoveries remains unexplored. Here we leveraged a randomized trial design, well-established threat-anticipation paradigm, and multidimensional battery of assessments to understand the consequences of 24-hour nicotine abstinence. The threat-anticipation paradigm had the expected consequences, amplifying subjective distress and arousal, and recruiting the canonical threat-anticipation network. Abstinence increased smoking urges and withdrawal symptoms, and potentiated threat-evoked distress, but had negligible consequences for EAc threat reactivity, raising questions about the translational relevance of prominent animal and human models of addiction. These observations provide a framework for conceptualizing nicotine abstinence and withdrawal, with implications for basic, translational, and clinical science., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Kim et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Viral lysis of phytoplankton is one of the most common forms of death on Earth. Building on an assay used extensively to assess rates of phytoplankton loss to predation by grazers, lysis rates are increasingly quantified through dilution-based techniques. In this approach, dilution of viruses and hosts are expected to reduce infection rates and thus increase host net growth rates (i.e., accumulation rates). The difference between diluted and undiluted host growth rates is interpreted as a measurable proxy for the rate of viral lytic death. These assays are usually conducted in volumes ≥ 1 L. To increase throughput, we implemented a miniaturized, high-throughput, high-replication, flow cytometric microplate dilution assay to measure viral lysis in environmental samples sourced from a suburban pond and the North Atlantic Ocean. The most notable outcome we observed was a decline in phytoplankton densities that was exacerbated by dilution, instead of the increased growth rates expected from lowered virus-phytoplankton encounters. We sought to explain this counterintuitive outcome using theoretical, environmental, and experimental analyses. Our study shows that, while die-offs could be partly explained by a 'plate effect' due to small incubation volumes and cells adhering to walls, the declines in phytoplankton densities are not volume-dependent. Rather, they are driven by many density- and physiology-dependent effects of dilution on predation pressure, nutrient limitation, and growth, all of which violate the original assumptions of dilution assays. As these effects are volume-independent, these processes likely occur in all dilution assays that our analyses show to be remarkably sensitive to dilution-altered phytoplankton growth and insensitive to actual predation pressure. Incorporating altered growth as well as predation, we present a logical framework that categorizes locations by the relative dominance of these mechanisms, with general applicability to dilution-based assays., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Knowles et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

A key broadening participation challenge for science communication is to reach non-traditional audiences beyond those already interested in science. In this study we test a "Guerilla Science" approach that blends elements of access, by removing barriers to participation, with those of inclusion, by designing participant-centered activities, for reaching an art-interested adult audience at the FIGMENT art festival in New York City. Our results show that participants at Guerilla Science were similar to festival goers overall in their connection to and interest in science, indicating the effectiveness of the approach for engaging non-traditional audiences and those who might not self-select into science activities., Competing Interests: We have read the journal’s policy and the authors of this manuscript have the following competing interests: Mark Rosin is a part owner of Guerilla Science Global and sits on its Board of Directors. This does not alter our adherence to PLOS ONE policies on sharing data and materials. The remaining authors declare that there are no conflicts of interest, (Copyright: © 2023 Rosin et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Disturbances to forested watersheds often result in increases of nutrients and light to nearby streams. Such changes are generally expected to produce a shift to a more autotrophic aquatic ecosystem, with measurable increases in algae, and associated implications for food webs and fisheries. Although this paradigm is widely established, results from our 10-year study (2007-2016) in 12 headwater streams and four sites downstream in the Trask River Watershed (Oregon, USA), did not concur. In 2012, one watershed was thinned, three were clearcut harvested with variable buffers and three with uniform riparian buffers. After harvest, light to the stream surface significantly increased at the three watersheds with variable buffers while dissolved inorganic nitrogen (DIN) significantly increased in all of the clearcut harvested streams. Despite the increase in DIN and light, algal standing stocks and chlorophyll a concentrations did not significantly increase. The common assumption of increased autotrophic responses in stream food webs following increases of nitrogen and light was not supported here. We postulate the co-limitation of nutrients, driven by low phosphorus concentrations, which unlike DIN did not increase post-harvest, and the characteristics of the algal community, which were dominated by low light adapted diatoms rather than green algae, contributed to our findings of no responses for standing stocks of epilithic algae or concentrations of chlorophyll a. The inclusion of multiple statistical analyses provided more certainty around our findings. This study documents responses to current forest practices and provides cautionary information for management and restoration activities aiming to increase fish abundance and standing stocks by opening riparian canopies and adding nutrients., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

DNA metabarcoding of pollen is a useful tool for studying bee foraging ecology. However, several questions about this method remain unresolved, including the extent to which sequence read data is quantitative, which type of sequence count removal threshold to use and how that choice affects our ability to detect rare flower visits, and how sequence artefacts may confound conclusions about bee foraging behavior. To address these questions, we isolated pollen from five plant species and created treatments comprised of pollen from each species alone and combinations of pollen from multiple species that varied in richness and evenness. We used ITS2 and rbcL metabarcoding to identify plant species in the samples, compared the proportion of pollen by mass to the proportion of sequencing reads for each plant species in each treatment, and analyzed the sequencing data using both liberal and conservative thresholds. We collected pollen from foraging bees, analyzed metabarcoding data from those samples using each threshold, and compared the differences in the pollinator networks constructed from the data. Regardless of the threshold used, the relationship between the proportion of pollen by mass and sequencing reads was inconsistent, suggesting that the number of sequence reads is a poor proxy for pollen abundance in mixed-species samples. Using a liberal threshold resulted in greater detection of original plant species in mixtures but also detected additional species in mixtures and single-species samples. The conservative threshold reduced the number of additional plant species detected, but several species in mixtures were not detected above the threshold, resulting in false negatives. Pollinator networks produced using the two thresholds differed and illustrated tradeoffs between detection of rare species and estimation of network complexity. Threshold selection can have a major effect on conclusions drawn from studies using metabarcoding of bee pollen to study plant-pollinator interactions., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

As contemporary wildfire activity intensifies across the western United States, there is increasing recognition that a variety of forest management activities are necessary to restore ecosystem function and reduce wildfire hazard in dry forests. However, the pace and scale of current, active forest management is insufficient to address restoration needs. Managed wildfire and landscape-scale prescribed burns hold potential to achieve broad-scale goals but may not achieve desired outcomes where fire severity is too high or too low. To explore the potential for fire alone to restore dry forests, we developed a novel method to predict the range of fire severities most likely to restore historical forest basal area, density, and species composition in forests across eastern Oregon. First, we developed probabilistic tree mortality models for 24 species based on tree characteristics and remotely sensed fire severity from burned field plots. We applied these estimates to unburned stands in four national forests to predict post-fire conditions using multi-scale modeling in a Monte Carlo framework. We compared these results to historical reconstructions to identify fire severities with the highest restoration potential. Generally, we found basal area and density targets could be achieved by a relatively narrow range of moderate-severity fire (roughly 365-560 RdNBR). However, single fire events did not restore species composition in forests that were historically maintained by frequent, low-severity fire. Restorative fire severity ranges for stand basal area and density were strikingly similar for ponderosa pine (Pinus ponderosa) and dry mixed-conifer forests across a broad geographic range, in part due to relatively high fire tolerance of large grand (Abies grandis) and white fir (Abies concolor). Our results suggest historical forest conditions created by recurrent fire are not readily restored by single fires and landscapes have likely passed thresholds that preclude the effectiveness of managed wildfire alone as a restoration tool., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

For centuries, humans occupying arid regions of North America have maintained an intricate relationship with Agave (Agavoideae, Asparagaceae). Today Agave cultivation, primarily for beverage production, provides an economic engine for rural communities throughout Mexico. Among known dryland-farming methods, the use of rock piles and cattle-grazed areas stand out as promising approaches for Agave cultivation. Identifying new cultivation areas to apply these approaches in Arizona, USA and Sonora, Mexico warrants a geographic assessment of areas outside the known ranges of rock piles and grasslands. The objective of this study was to predict areas for dryland-farming of Agave and develop models to identify potential areas for Agave cultivation. We used maximum entropy (MaxEnt) ecological-niche-modeling algorithms to predict suitable areas for Agave dryland farming. The model was parameterized using occurrence records of Hohokam rock piles in Arizona and grassland fields cultivated with Agave in Sonora. Ten environmental-predictor variables were used in the model, downloaded from the WorldClim 2 climate database. The model identified potential locations for using rock piles as dryland-farming methods from south-central Arizona to northwestern Sonora. The Agave-grassland model indicated that regions from central to southern Sonora have the highest potential for cultivation of Agave, particularly for the species Agave angustifolia. Results suggest that there are many suitable areas where rock piles can be used to cultivate Agave in the Sonoran Desert, particularly in the border of southeastern Arizona and northwest Sonora. Likewise, cattle-grazing grasslands provide a viable environment for cultivating Agave in southern Sonora, where the expanding bacanora-beverage industry continues to grow and where different Agave products (e.g., syrups, fructans, saponins, and medicinal compounds) can potentially strengthen local economies., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Ortiz Cano et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)