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All sourcesNASA NTRSDOE OSTI
DOE OSTI2025

2024 Results for Avian Monitoring at the Technical Area 36 Minie Site, Technical Area 39 Point 6, Technical Area 16 Burn Ground, and DARHT at Los Alamos National Laboratory

Los Alamos National Laboratory (LANL) biological subject matter experts in the Environmental Protection and Compliance Division initiated a multi-year program in 2013 to monitor avifauna (birds) at two open detonation sites and one open burn site on LANL property. Additional monitoring began in 2017 at a third firing site, the Dual-Axis Radiographic Hydrodynamic Test (DARHT) Facility. In this annual report, we compare monitoring results from these efforts among years to identify and evaluate firing and open burn site impacts on the local bird community. The objectives of this study are • to determine whether LANL operations impact bird abundance, species richness, or diversity; • to examine occupancy and nest success of secondary-cavity nesting birds that use nest boxes; and • to examine chemical concentrations (such as radionuclides, inorganic elements, and/or organic compounds) in nonviable eggs and deceased nestlings that are collected opportunistically with the upper-level bounds of background concentrations, when available. During May through July 2024, LANL biologists completed multiple avian point count surveys at each of the following treatment sites: • Technical Area (TA) 36 Minie Site, • TA-39 Point 6, • TA-16 Burn Ground, and • DARHT. We recorded a total of 1,088 birds that represented 65 species at the four treatment sites and compared these results with data from their associated control sites. In 2024, abundance and species richness at treatment and control sites continued to trend similarly from year to year, with minor random deviations expected from bird communities. Species richness at firing sites differed little from the previous year’s values. Two new bird species were observed at the firing sites—cedar waxwing (Bombycilla cedrorum) and pinyon jay (Gymnorhinus cyanocephalus). Shannon diversity values at TA-36 Minie Site, TA-39, and DARHT were statistically higher than one or more of their associated controls. Annual species diversity at treatment sites was high in 2024 across all firing sites relative to similar habitat control sites. We also monitored avian nest boxes to compare occupancy and nest success data from nest boxes at treatment sites with the overall avian nest box monitoring network and against a subset of relevant control sites. Nest box success has decreased at both treatment and control sites since monitoring began, suggesting that overlapping climatic factors are responsible for patterns of declining nest success. In 2024, nonviable avian eggs and one nestling were opportunistically collected at Bandelier National Monument, TA-16 Burn Ground, TA-36 Minie, TA-39 Point 6, and DARHT. All egg samples and the one nestling sample were evaluated for per- and polyfluoroalkyl substances, which were detected from all locations, including the control site at Bandelier National Monument. Overall results from 2024 continue to suggest that operations at the four treatment sites are not negatively impacting bird populations. This long-term project will continue to monitor for any changes over time.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2022

Comparison and validation of the QuEChERSER mega-method for determination of per- and polyfluoroalkyl substances in foods by liquid chromatography with high-resolution and triple quadrupole mass spectrometry

Instances of food contamination with per- and polyfluoroalkyl substances (PFAS) continue to occur globally, but sample preparation and analytical methods are quite limited and often monitor for a small percentage of known PFAS. This study aimed to evaluate, validate, and compare performance of two instruments with the recently developed “quick, easy, cheap, effective, rugged, safe, efficient, and robust” (QuEChERSER) sample preparation mega-method – a method developed to monitor chemicals over a broad range of physicochemical properties. Initial evaluation of the QuEChERSER mega-method for determination of PFAS in food demonstrated recoveries, matrix interferences, and co-extractive removal comparable to (or better than) US Food and Drug Administration (FDA) and USDA Food Safety and Inspection Service (FSIS) methods. Subsequent validation of QuEChERSER in beef, catfish, chicken, pork, liquid eggs, and powdered eggs on a high-resolution mass spectrometer achieved acceptable recoveries (70–120%) and precision (RSDs ≤20%) for all 33 target analytes at the 1 and 5 ng g –1 levels and 67–88% of analytes at the 0.1 ng g –1 level, depending on the matrix. Additional validation was performed by tandem mass spectrometry on a triple quadrupole instrument. This approach provided no non-detects and better recoveries at the 0.1 ng g –1 level than the HRMS method but exhibited more variability at 1 and 5 ng g –1 spiking levels. Analysis of NIST SRMs 1946 and 1947 gave accuracies of 70–117%. Furthermore, these results demonstrate the capability of combining PFAS analysis with a mega-method previously validated for 350 analytes, while collecting non-target data for future retrospective analysis of emerging alternatives with a high-resolution mass spectrometry method.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2020

Directed energy system technology for the control of soilborne fungal pathogens and plant‐parasitic nematodes

Abstract BACKGROUND It is challenging to manage soilborne pathogens and plant‐parasitic nematodes using sustainable practices. Here, we evaluated a novel energy application system, Directed Energy System (DES). This system generates pulses of energy capable of impacting selected biological organisms. The oomycete Phytophthora cinnamomi , the fungus Verticillium dahliae , and the plant‐parasitic nematodes Meloidogyne hapla and Globodera ellingtonae were added to soil. Then DES‐generated energy was applied to soil and impacts on target organisms were determined. RESULTS DES applied at 20, 40 and 70 J cm −3 to P. cinnamomi and V. dahliae resulted in ≥50% and 92% reductions (respectively) of propagules per gram of soil in comparison to the untreated control. There was a significant reduction of M. hapla eggs per gram of host tomato root between the untreated control, and 2.2, 13 and 25 J cm −3 DES dosages applied pre‐ or post‐planting. Additionally, an 84% reduction in hatch from G. ellingtonae encysted eggs after treatment with 70 J cm −3 DES was observed. The dosages ranged from 40 or 80V mm −1 for nematodes to 200 V mm −1 for fungi. CONCLUSION DES‐generated energy reduced survival of the soilborne pathogens P. cinnamomi and V. dahlia, and the plant‐parasitic nematodes M. hapla and G. ellingtonae . The application of this technology to a field setting remains to be considered. Published 2020. This article is a U.S. Government work and is in the public domain in the USA. Pest Management Science published by Wiley Periodicals, Inc. on behalf of © 2020 Society of Chemical Industry.

Riga, Ekaterini↗
DOE OSTI2025

Persistence of bacterial-mediated anti-rotifer protection in preliminary outdoor cultivation trial for Microchloropsis salina

Outdoor algal cultivation systems are susceptible to a wide variety of deleterious species. In previously published studies, we observed protection using microbial consortia at laboratory scale cultures; Microchloropsis salina in the presence of microbial consortia were protected from grazing from the marine rotifer, Brachionus plicatilis. Our objective for the present work was to determine if this protection conferred by microbial consortia in controlled laboratory experiments would persist in an open, outdoor multi-liter cultivation system. We found that algal protection did persist as evidenced by the presence of fewer motile rotifers and decreased rotifer-associated egg counts for the consortia-treated outdoor cultures. Due to the low temperature and light conditions that reduced growth of the algae outdoors, we performed an indoor laboratory assay which also confirmed the persistence of algal protection. Lastly, the lower numbers of motile rotifers and fewer rotifer-associated eggs in the consortia-treated algal cultures suggests a possible protective mechanism by the consortia through interfering with the rotifer lifecycle or reproduction. Finally, these initial results support the possibility that low cost, prophylactic treatments with microbial consortia can protect algae from deleterious species in outdoor cultivation systems.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

Quantifying agricultural productive use of energy load in Sub-Saharan Africa and its impact on microgrid configurations and costs

The use of advanced energy technologies for agricultural purposes—such as irrigation, refrigeration, crop processing, and egg incubation—has the potential to increase crop yield, reduce vulnerability to changing precipitation patterns, increase shelf life, strengthen income and employment opportunities in rural areas, and reduce emissions by displacing fossil fuel-based technologies. These productive uses of energy (PUE) in remote areas could potentially be powered by microgrids that additionally serve otherwise unelectrified communities, most of which are located in rural Sub-Saharan Africa. Here, in this paper, we use high-resolution geospatial data to estimate the end-use electricity demand for a range of agricultural PUE across Sub-Saharan Africa, and we share these data in an open-access mapping tool. Next, we use REopt®, a techno-economic optimization model of energy systems, to determine the cost and system sizing implications of incorporating agricultural PUE into microgrid designs in Kenya and Zambia. We estimate the upper bound of agricultural PUE demand for irrigation, milling, shelling, refrigeration, and egg incubation across Sub-Saharan to be 16.8 TWh/yr. We find that incorporating local agricultural PUE into microgrid system designs increases the required system sizing while having minimal impact on the levelized cost of energy of these systems. Our analysis is the first to demonstrate the PUE potential in the agricultural sector at a 10x10-kilometer resolution across Sub-Saharan Africa and to show, at scale, how site-specific PUE can impact the cost and sizing of microgrids that are otherwise deployed to serve local household and community load.

24 POWER TRANSMISSION AND DISTRIBUTION↗
DOE OSTI2021

Metal Ion Fluxes Controlling Amphibian Fertilization

Mammalian oocytes undergo major changes in zinc content and localization to be fertilized, the most striking being the rapid exocytosis of over 10 billion zinc ions in what are known as zinc sparks. Here, we report that fertilization of amphibian Xenopus laevis eggs also initiates a zinc spark that progresses across the cell surface in coordination with dynamic calcium waves. This zinc exocytosis is accompanied by a newly recognized loss of intracellular manganese. Synchrotron-based X-ray fluorescence and analytical electron microscopy reveal that zinc and manganese are sequestered in a system of cortical granules that are abundant at the animal pole. Through electron-nuclear double-resonance studies, we rule out Mn2+ complexation with phosphate or nitrogenous ligands in intact eggs, but the data are consistent with a carboxylate coordination environment. Our observations suggest that zinc and manganese fluxes are a conserved feature of fertilization in vertebrates and that they function as part of a physiological block to polyspermy.

Seeler, John F.↗
DOE OSTI2024

Improving the precision of forces in real-space pseudopotential density functional theory

The high-order finite difference real-space pseudopotential density functional theory (DFT) approach is a valuable method for large-scale, massively parallel DFT calculations. A significant challenge in the approach is the oscillating “egg-box” error introduced by aliasing associated with a coarse grid spacing. To address this issue while minimizing computational cost, we developed a finite difference interpolation (FDI) scheme [Roller et al., J. Chem. Theory Comput. 19, 3889 (2023)] as a means of exploiting the high resolution of the pseudopotential to reduce egg-box effects systematically. Here, we show an implementation of this method in the PARSEC code and examine the practical utility of the combination of FDI with additional methods for improving force precision and/or reducing its computational cost, including orbital-based forces, compensating charges (namely, adding and subtracting a judiciously chosen charge density such that the total density is unaltered), and a modified spatial domain in which the real-space grid is defined. Using selected small molecules, as well as metallic Li, as test cases, we show that a combination of all four aspects leads to a significant reduction in computational cost while retaining a high level of precision that supports accurate structures and vibrational spectra, as well as stable and accurate molecular dynamics runs.

Chemistry↗
DOE OSTI2022

Survival estimates across five life stages of redfin ( Perca fluviatilis ) exposed to simulated pumped-storage hydropower stressors

The global prevalence of pumped-storage hydropower (PSH) is expected to grow exponentially as countries transition to renewable energy sources. Compared to conventional hydropower, little is currently known regarding PSH impacts on aquatic biota. This study estimated the survival of five life stages (egg, two larval stages, juvenile and adult) of redfin (European) perch (Perca fluviatilis) following passage through a PSH facility during the pumping phase. This was achieved by simulating the individual stressors expected to occur during passage through a 2000-MW PSH facility using laboratory-simulated (shear strain and extreme compression) and modelling (blade strike, BS) approaches. Our results indicate that redfin could survive the shear, pressure and BS stressors expected within the PSH facility, but impacts varied among life stages. Juvenile survival was >70% across all shear strain rates, while the survival of eggs and larvae declined markedly as strain rate increased. All life stages had high survival when exposed to rapid compression and BS. The high survival of redfin to the stressors tested suggests the PSH facility could facilitate the passage of redfin during the pumping phase from the lower to the higher elevation reservoir. This outcome would be welcomed in situations where the species is native, but could have adverse implications for the conservation of native biota where the species is considered a pest.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2025

Impact of developmental methylmercury exposure on avian embryonic development, hatchling growth, and survival

Abstract Methylmercury (MeHg) is a globally ubiquitous and persistent environmental toxicant that negatively affects animal behavior, health, and reproduction. In birds, MeHg is transferred from female to egg, contaminating embryos during sensitive stages of early embryonic development and growth. This toxic exposure in the prenatal environment not only induces mortality but also possible lasting impacts on physiology, health, and survival, even once hatched. The purpose of our study was to further elucidate the negative effects of MeHg exposure during avian embryonic development and explore how such exposure can impact offspring development, growth, and survival, both in ovo and posthatch. To assess this, we experimentally dosed fertile mallard and wood duck eggs with MeHg II chloride and reared developing embryos and ducklings to various endpoints. We found that embryos not only readily accumulated MeHg throughout incubation, but they also displayed varying dose-dependent disparities in body mass and morphometrics, with control individuals being larger during early and late embryonic stages of development (p < 0.05). Furthermore, hatched offspring exposed to MeHg exhibited increasingly slower growth rates between 7 to 10 and 10 to 15 days posthatch (p < 0.05), and were found to have lower survival probabilities both under controlled laboratory conditions (p < 0.005), and in the natural environment (p < 0.05). Our findings on the detrimental effects of MeHg on avian embryos and hatchlings emphasize the need for more proactive means of environmental protection and remediation to protect vulnerable avian populations and the ecosystems they inhabit.

Leaphart, James C.↗
DOE OSTI2020

Incubation Temperature Affects Duckling Body Size and Food Consumption Despite No Effect on Associated Feeding Behaviors

Synopsis Developmental conditions can have consequences for offspring fitness. For example, small changes (<1°C) in average avian incubation temperature have large effects on important post-hatch offspring phenotypes, including growth rate, thermoregulation, and behavior. Furthermore, average incubation temperatures differ among eggs within the same nest, to the extent (i.e., >1°C) that differences in offspring phenotypes within broods should result. A potential consequence of within-nest incubation temperature variation is inequality in behaviors that could cause differences in resource acquisition within broods. To investigate this, we incubated wood duck (Aix sponsa) eggs at one of two ecologically-relevant incubation temperatures (35°C or 36°C), formed mixed-incubation temperature broods after ducklings hatched, and conducted trials to measure duckling behaviors associated with acquisition of heat (one trial) or food (three trials). Contrary to our predictions, we found no effect of incubation temperature on duckling behaviors (e.g., time spent occupying heat source, frequency of feeding bouts). However, we found evidence that ducklings incubated at the higher temperature consumed more food during the 1-h feeding trials, and grew faster in body mass and structural size (culmen and tarsus) throughout the study, than those incubated at the lower temperature. Apparent food consumption during the trials was positively related to culmen length, suggesting that differences in food consumption may be driven by structural size. This could result in positive feedback, which would amplify size differences between offspring incubated at different temperatures. Thus, our study identifies incubation temperature as a mechanism by which fitness-related phenotypic differences can be generated and even amplified within avian broods.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Ethyl Formate as a Methyl Bromide Alternative for Fumigation of Citrus: Efficacy, Fruit Quality, and Workplace Safety

Abstract Ethyl formate (EF) was evaluated as a potential alternative to methyl bromide (MB) for phytosanitary treatment of imported citrus fruit in the Republic of Korea. Planococcus citri (Risso) (Hemiptera: Pseudococcidae), a mealybug with known tolerance against EF and MB, was used as a representative pest to test efficacy of the two fumigants against eggs. In nine commercial-scale refrigerated container (67.5 m3) trials using imported orange, lemon and grapefruit, EF applied at the currently approved dose for citrus (70 g·m-3 at 5°C for 4 h, developed for Aspidiotus excisus Green (Hemiptera: Diaspididae), a species less EF tolerant than P. citri) resulted in 76.9–98.3% mortality of P. citri eggs. The EF treatment did not affect the sugar content or the color of peel and pulp of the treated fruit. When oranges were treated according to the current MB (64 g·m-3 at >5°C for 2 h) or EF treatment guidelines, the concentration of fumigant around the fruit fluctuated between 9.4 and 185.1 ppm for EF and 9.5–203.0 ppm for MB during the 72-h post-fumigation processes (venting [0–2 h], transportation to storage [2–24 h], and storage periods [24–72 h]) with both EF and MB maintained between 10 and 100 ppm during the storage period. Considering the efficacy of EF, its apparent lack of phytotoxicity, and its more manageable threshold limit value for humans (100 ppm EF compared to 1 ppm MB for an 8-h time weighted average exposure), our results suggest that EF may be a promising alternative to MB for the phytosanitary treatment of imported citrus in Korea.

Entomology↗
DOE OSTI2022

Generating Ectropis obliqua axenic larvae in support of functional microbiome studies

Abstract Ectropis obliqua Prout (Lepidoptera: Geometridae) is a serious leaf‐eating pest in tea gardens throughout China, with rich bacterial communities in the larval gut. Few studies have investigated whether these bacteria play an important role in growth, development, and adaptability of E. obliqua . Axenic larval models are a useful tool for the study of interactions between gut bacteria and host insects. At present, artificial diet and antibiotics are typically used to remove larval gut bacteria for study; however, this method may change the natural food of insects and the habitat provided by their host plants, and cannot completely eliminate insect gut bacteria. Therefore, in order to establish an axenic insect feeding model, we first investigated the effects of egg age, different disinfectant concentration, and treatment time on the eggs of E. obliqua through orthogonal experiments. Next, we fed E. obliqua larvae tissue‐cultured tea seedlings under aseptic conditions. Thus, we successfully generated a feeding model of gut axenic larvae in the laboratory. Based on it, we then compared the mortality of axenic and non‐axenic E. obliqua larvae infected with the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuill. These results demonstrated the feasibility of this axenic larval model and that B. bassiana is a broad‐spectrum entomogenous fungus with a toxic effect on E. obliqua .

Su, Qingqing↗
DOE OSTI2023

Scoping review of Culex mosquito life history trait heterogeneity in response to temperature

Abstract Background Mosquitoes in the genus Culex are primary vectors in the US for West Nile virus (WNV) and other arboviruses. Climatic drivers such as temperature have differential effects on species-specific changes in mosquito range, distribution, and abundance, posing challenges for population modeling, disease forecasting, and subsequent public health decisions. Understanding these differences in underlying biological dynamics is crucial in the face of climate change. Methods We collected empirical data on thermal response for immature development rate, egg viability, oviposition, survival to adulthood, and adult lifespan for Culex pipiens, Cx. quinquefasciatus, Cx. tarsalis , and Cx. restuans from existing literature according to the PRISMA scoping review guidelines. Results We observed linear relationships with temperature for development rate and lifespan, and nonlinear relationships for survival and egg viability, with underlying variation between species. Optimal ranges and critical minima and maxima also appeared varied. To illustrate how model output can change with experimental input data from individual Culex species, we applied a modified equation for temperature-dependent mosquito type reproduction number for endemic spread of WNV among mosquitoes and observed different effects. Conclusions Current models often input theoretical parameters estimated from a single vector species; we show the need to implement the real-world heterogeneity in thermal response between species and present a useful data resource for researchers working toward that goal. Graphical Abstract

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Assessing the Ability of Developmentally Precocious Estrogen Signaling to Recapitulate Ovarian Transcriptomes and Follicle Dynamics in Alligators from a Contaminated Lake

Concern has grown in recent decades over anthropogenic contaminants that interfere with the functioning of endocrine hormones. However, mechanisms connecting developmental processes to pathologies associated with endocrine-disrupting chemical (EDC) exposure are poorly understood in naturally exposed populations. We sought to a) characterize divergence in ovarian transcriptomic and follicular profiles between alligators originating from a historically EDC-contaminated site, Lake Apopka, and a reference site; b) test the ability of developmentally precocious estrogen exposure to recapitulate site-associated patterns of divergence; and c) test whether treatment with exogenous follicle-stimulating hormone (FSH) is capable of rescuing phenotypes associated with contaminant exposure and/or embryonic estrogen treatment. Alligators eggs were collected from a contaminated site and a reference site, and a subset of eggs from the reference site were treated with estradiol (E2) during embryonic development prior to gonadal differentiation. After hatching, alligators were raised under controlled laboratory settings for 5 months. Juveniles from both sites were divided and treated with exogenous FSH. Histological analyses and RNA-sequencing were conducted to characterize divergence in ovarian follicle dynamics and transcriptomes between sites, between reference and E2-treated animals, and between FSH-treated and nontreated animals. We observed broad site-of-origin divergence in ovarian transcriptomes and reductions in ovarian follicle density between juvenile alligators from Lake Apopka and the reference site. Treating embryos from the reference site with E2 overwhelmingly recapitulated transcriptional and histological alterations observed in Lake Apopka juveniles. Ovarian phenotypes observed in Lake Apopka alligators or resulting from estrogen treatment were only partially rescued by treatment with exogenous FSH. Recapitulation of ovarian abnormalities by precocious E2 revealed a relatively simple mechanism underlying contaminant-induced pathologies in a historical example of environmental endocrine disruption. Findings reported here support a model where the developmental timing of estrogen signaling has the potential to permanently alter ovarian organization and function.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

A new class of antibodies that overcomes a steric barrier to cross-group neutralization of influenza viruses

Antibody titers that inhibit the influenza virus hemagglutinin (HA) from engaging its receptor are the accepted correlate of protection from infection. Many potent antibodies with broad, intra-subtype specificity bind HA at the receptor binding site (RBS). One barrier to broad H1-H3 cross-subtype neutralization is an insertion (133a) between positions 133 and 134 on the rim of the H1 HA RBS. We describe here a class of antibodies that overcomes this barrier. These genetically unrestricted antibodies are abundant in the human B cell memory compartment. Analysis of the affinities of selected members of this class for historical H1 and H3 isolates suggest that they were elicited by H3 exposure and broadened or diverted by later exposure(s) to H1 HA. RBS mutations in egg-adapted vaccine strains cause the new H1 specificity of these antibodies to depend on the egg adaptation. The results suggest that suitable immunogens might elicit 133a-independent, H1-H3 cross neutralization by RBS-directed antibodies.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2025

Multi-contrast machine learning improves schistosomiasis diagnostic performance

Schistosomiasis currently affects over 250 million people and remains a public health burden despite ongoing global control efforts. Conventional microscopy is a practical tool for diagnosis and screening ofSchistosoma haematobium, but identification of eggs requires a skilled microscopist. Here we present a machine learning (ML)-based strategy for automated detection ofS. haematobiumthat combines two imaging contrasts, brightfield (BF) and darkfield (DF), to improve diagnostic performance. We collected BF and DF images of urine samples, many of them containingS. haematobiumeggs, during two different field studies in Côte d’Ivoire using a mobile phone-based microscope, the SchistoScope. We then trained separate egg-detection ML models and compared the patient-level performance of BF and DF models alone to combinations of BF and DF models, using annotations from trained microscopists as the gold standard. We found that models trained on DF images, and almost all BF and DF combinations, performed significantly better than models trained on BF images only. When models were trained on images from the first field study (n = 349 patients, 748 images of each contrast), patient-level classification performance on patient images from the second study (n = 375 patients, 752 images of each contrast) met the WHO Diagnostic Target Product Profile (TPP) sensitivity and specificity for the monitoring and evaluation use case (sensitivity for all models and combinations was >75% when evaluated at a confidence score threshold that resulted in specificity >96.5%). When we used images from both field studies for the training set, performance of the models was improved. Overall, this work shows that the use of DF and BF increases the performance of ML models on images from devices with low-cost optics, while retaining the portability, power, and time-to-results of the WHO’s diagnostic TPP. DF requires no additional sample preparation and does not increase the complexity of the imaging system. It thus offers a practical means to improve performance of automated diagnostics forS. haematobiumas well as other microscopy-based diagnostics.

Infectious Diseases↗
DOE OSTI2020

Leisingera sp. ANG59 & Pseudoalteromonas sp. JC28 Genomes Overview

The Hawaiian bobtail squid, Euprymna scolopes, has a symbiotic bacterial consortium in the accessory nidamental gland (ANG), a female reproductive organ that protects eggs against fouling microorganisms. To test the antibacterial activity of ANG community members, 19 bacterial isolates were screened for their ability to inhibit Gram-negative and Gram-positive bacteria, of which two strains were inhibitory. These two antibacterial isolates, Leisingera sp. ANG59 and Pseudoalteromonas sp. JC28, were subjected to further genomic characterization. Genomic analysis of Leisingera sp. ANG59 revealed a biosynthetic gene cluster encoding the antimicrobial compound indigoidine. The genome of Pseudoalteromonas sp. JC28 had a 14-gene cluster with >95% amino acid identity to a known bromoalterochromide (BAC) cluster. Chemical analysis confirmed production of known BACs, BAC-A/A′ (compounds 1a/1b), as well as two new derivatives, BAC-D/D′ (compounds 2a/2b). Extensive nuclear magnetic resonance (NMR) analyses allowed complete structural elucidation of compounds 2a/2b, and the absolute stereochemistry was unambiguously determined using an optimized Marfey’s method. The BACs were then investigated for in vitro antibacterial, antifungal, and nitric oxide (NO) inhibitory activity. Compounds 1a/1b were active against the marine bacteria Bacillus algicola and Vibrio fischeri, while compounds 2a/2b were active only against B. algicola. Compounds 1a/1b inhibited NO production via lipopolysaccharide (LPS)-induced inflammation in RAW264.7 macrophage cells and also inhibited the pathogenic fungus Fusarium keratoplasticum, which, coupled with their antibacterial activity, suggests that these polyketide-nonribosomal peptides may be used for squid egg defense against potential pathogens and/or fouling microorganisms. These results indicate that BACs may provide Pseudoalteromonas sp. JC28 an ecological niche, facilitating competition against nonsymbiotic microorganisms in the host’s environment.​

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

The influence of incubation temperature on offspring traits varies across northern and southern populations of the American alligator ( Alligator mississippiensis )

Maternal provisioning and the developmental environment are fundamental determinants of offspring traits, particularly in oviparous species. However, the extent to which embryonic responses to these factors differ across populations to drive phenotypic variation is not well understood. Here, we examine the contributions of maternal provisioning and incubation temperature to hatchling morphological and metabolic traits across four populations of the American alligator (Alligator mississippiensis), encompassing a large portion of the species' latitudinal range. Our results show that whereas the influence of egg mass is generally consistent across populations, responses to incubation temperature show population-level variation in several traits, including mass, head length, head width, and residual yolk mass. Additionally, the influence of incubation temperature on developmental rate is greater at northern populations, while the allocation of maternal resources toward fat body mass is greater at southern populations. Overall, our results suggest that responses to incubation temperature, relative to maternal provisioning, are a larger source of interpopulation phenotypic variation and may contribute to the local adaptation of populations.

59 BASIC BIOLOGICAL SCIENCES↗