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At least 739 records · Page 41

Phylodynamics of SARS-CoV-2 Lineages B.1.1.7, B.1.1.529 and B.1.617.2 in Nigeria Suggests Divergent Evolutionary Trajectories

Background: The early months of the COVID-19 pandemic were characterized by high transmission rates and mortality, compounded by the emergence of multiple SARS-CoV-2 lineages, including Variants of Concern (VOCs). This study investigates the phylodynamic and spatio-temporal trends of VOCs during the peak of the pandemic in Nigeria. Methods: Whole-genome sequencing (WGS) data from three major VOCs circulating in Nigeria, B.1.1.7 (Alpha), B.1.617.2 (Delta), and B.1.1.529 (Omicron), were analyzed using tools such as Nextclade, R Studio v 4.2.3, and BEAST X v 10.5.0. The spatial distribution, evolutionary history, viral ancestral introductions, and geographic dispersal patterns were characterized. Results: Three major lineages following WHO nomenclature were identified: Alpha, Delta, and Omicron. The Delta variant exhibited the widest geographic spread, detected in 14 states, while the Alpha variant was the least distributed, identified in only eight states but present across most epidemiological weeks studied. Evolutionary rates varied slightly, with Alpha exhibiting the slowest rate (2.66 × 10 −4 substitutions/site/year). Viral population analyses showed distinct patterns: Omicron sustained elevated population growth over time, while Delta declined after initial expansion. The earliest Times to Most Recent Common Ancestor (TMRCA) were consistent with the earliest outbreaks of SARS-CoV-2 globally. Geographic transmission analysis indicated a predominant coastal-to-inland spread for all variants, with Omicron showing the most diffuse dispersal, highlighting commercial routes as significant drivers of viral diffusion. Conclusion: The SARS-CoV-2 epidemic in Nigeria was characterized by multiple variant introductions and a dominant coastal-to-inland spread, emphasizing that despite lockdown measures, commercial trade routes played a critical role in viral dissemination. These findings provide insights into pandemic control strategies and future outbreak preparedness.

Nigeria↗

Deep Learning for Automated Detection and Identification of Migrating American Eel Anguilla rostrata from Imaging Sonar Data

Adult American eels (Anguilla rostrata) are vulnerable to hydropower turbine mortality during outmigration from growth habitat in inland waters to the ocean where they spawn. Imaging sonar is a reliable and proven technology for monitoring of fish passage and migration; however, there is no efficient automated method for eel detection. We designed a deep learning model for automated detection of adult American eels from sonar data. The method employs convolution neural network (CNN) to distinguish between 14 images of eels and non-eel objects. Prior to image classification with CNN, background subtraction and wavelet denoising were applied to enhance sonar images. The CNN model was first trained and tested on data obtained from a laboratory experiment, which yielded overall accuracies of >98% for image-based classification. Then, the model was trained and tested on field data that were obtained near the Iroquois Dam located on the St. Lawrence River; the accuracy achieved was commensurate with that of human experts.

13 HYDRO ENERGY↗

Remote Sensing Detection of Growing Season Freeze-Induced Defoliation of Montane Quaking Aspen (Populus tremuloides) in Southern Utah, USA

Growing season freeze events pose a threat to quaking aspen (Populus tremuloides Michx.), leading to canopy defoliation, reduced vigor, and increased mortality, especially for declining montane populations western North America. Detecting the spatial distribution and progression of this damage is challenging due to limited in situ observations in this region. This study represents the first attempt to comprehensively resolve the spatial extent of freeze-induced aspen canopy damage in southern Utah using multispectral remote sensing data. We developed an approach to detect the spatial and temporal dynamics of freeze-damaged aspen stands, focusing on a freeze event from 8–9 June 2020 in southern Utah. By integrating medium- (~250 to 500 m) and high-resolution (~10 m) satellite data, we employed the Normalized Difference Vegetation Index (NDVI) to compare post-freeze conditions with historical norms and pre-freeze conditions. Our analysis revealed NDVI reductions of 0.10 to 0.40 from pre-freeze values and a second flush recovery. We introduced a pixel-based method to evaluate freeze vulnerability, establishing a strong correlation (R values 0.78 to 0.82) between the onset of the first flush (NDVI > 0.50) and the accumulation of 100 growing degree days (GDD). These methods support the potential for retrospective assessments, proactive forest monitoring, and forecasting future risks.

54 ENVIRONMENTAL SCIENCES↗

Dose Response, Dosimetric, and Metabolic Evaluations of Replacement PFAS Perfluoro-(2,5,8-trimethyl-3,6,9-trioxadodecanoic) Acid (HFPO-TeA)

Few studies are available on the environmental and toxicological effects of perfluoroalkyl ether carboxylic acids (PFECAs), such as GenX, which are replacing legacy PFAS in manufacturing processes. To collect initial data on the toxicity and toxicokinetics of a longer-chain PFECA, male and female Sprague Dawley rats were exposed to perfluoro-(2,5,8-trimethyl-3,6,9-trioxadodecanoic) acid (HFPO-TeA) by oral gavage for five days over multiple dose levels (0.3–335.2 mg/kg/day). Clinically, we observed mortality at doses >17 mg/kg/day and body weight changes at doses ≤ 17 mg/kg/day. For the 17 mg/kg/day dose level, T3 and T4 thyroid hormone concentrations were significantly decreased (p < 0.05) from controls and HFPO-TeA plasma concentrations were significantly different between sexes. Non-targeted analysis of plasma and in vitro hepatocyte assay extractions revealed the presence of another GenX oligomer, perfluoro-(2,5-dimethyl-3,6-dioxanonanoic) acid (HFPO-TA). In vitro to in vivo extrapolation (IVIVE) parameterized with in vitro toxicokinetic data predicted steady-state blood concentrations that were within seven-fold of those observed in the in vivo study, demonstrating reasonable predictivity. The evidence of thyroid hormone dysregulation, sex-based differences in clinical results and dosimetry, and IVIVE predictions presented here suggest that the replacement PFECA HFPO-TeA induces a complex and toxic exposure response in rodents.

54 ENVIRONMENTAL SCIENCES↗

Mixture Effects of Per- and Polyfluoroalkyl Substances on Embryonic and Larval Sheepshead Minnows ( Cyprinodon variegatus )

Per- and polyfluoroalkyl substances (PFAS) are ubiquitous and persistent environmental contaminants originating from many everyday products. Perfluorooctane sulfonic acid (PFOS) and perfluorooctanoic acid (PFOA) are two PFAS that are commonly found at high concentrations in aquatic environments. Both chemicals have previously been shown to be toxic to fish, as well as having complex and largely uncharacterized mixture effects. However, limited information is available on marine and estuarine species. In this study, embryonic and larval sheepshead minnows (Cyprinodon variegatus) were exposed to several PFAS mixtures to assess lethal and sublethal effects. PFOS alone was acutely toxic to larvae, with a 96 h LC50 of 1.97 mg/L (1.64–2.16). PFOS + PFOA resulted in a larval LC 50 of 3.10 (2.62–3.79) mg/L, suggesting an antagonistic effect. These observations were supported by significant reductions in malondialdehyde (105% ± 3.25) and increases in reduced glutathione concentrations (43.8% ± 1.78) in PFOS + PFOA exposures compared to PFOS-only treatments, indicating reduced oxidative stress. While PFOA reduced PFOS-induced mortality (97.0% ± 3.03), perfluorohexanoic acid (PFHxA) and perfluorobutanoic acid (PFBA) did not. PFOS alone did not affect expression of peroxisome proliferator-activated receptor alpha (pparα) but significantly upregulated apolipoprotein A4 (apoa4) (112.4% ± 17.8), a downstream product of pparα, while none of the other individually tested PFAS affected apoa4 expression. These findings suggest that there are antagonistic interactions between PFOA and PFOS that may reduce mixture toxicity in larval sheepshead minnows through reduced oxidative stress. Elucidating mechanisms of toxicity and interactions between PFAS will aid environmental regulation and management of these ubiquitous pollutants.

59 BASIC BIOLOGICAL SCIENCES↗

A Novel Glutathione S-Transferase Gtt2 Class (VpGSTT2) Is Found in the Genome of the AHPND/EMS Vibrio parahaemolyticus Shrimp Pathogen

Glutathione S-transferases are a family of detoxifying enzymes that catalyze the conjugation of reduced glutathione (GSH) with different xenobiotic compounds using either Ser, Tyr, or Cys as a primary catalytic residue. We identified a novel GST in the genome of the shrimp pathogen V. parahaemolyticus FIM- S1708+, a bacterial strain associated with Acute Hepatopancreatic Necrosis Disease (AHPND)/Early Mortality Syndrome (EMS) in cultured shrimp. This new GST class was named Gtt2. It has an atypical catalytic mechanism in which a water molecule instead of Ser, Tyr, or Cys activates the sulfhydryl group of GSH. The biochemical properties of Gtt2 from Vibrio parahaemolyticus (VpGSTT2) were characterized using kinetic and crystallographic methods. Recombinant VpGSTT2 was enzymatically active using GSH and CDNB as substrates, with a specific activity of 5.7 units/mg. Low affinity for substrates was demonstrated using both Michaelis–Menten kinetics and isothermal titration calorimetry. The crystal structure showed a canonical two-domain structure comprising a glutathione binding G-domain and a hydrophobic ligand H domain. A water molecule was hydrogen-bonded to residues Thr9 and Ser 11, as reported for the yeast Gtt2, suggesting a primary role in the reaction. Molecular docking showed that GSH could bind at the G-site in the vicinity of Ser11. G-site mutationsT9A and S11A were analyzed. S11A retained 30% activity, while T9A/S11A showed no detectable activity. VpGSTT2 was the first bacterial Gtt2 characterized, in which residues Ser11 and Thr9 coordinated a water molecule as part of a catalytic mechanism that was characteristic of yeast GTT2. The GTT2 family has been shown to provide protection against metal toxicity; in some cases, excess heavy metals appear in shrimp ponds presenting AHPND/EMS. Further studies may address whether GTT2 in V. parahaemolyticus pathogenic strains may provide a competitive advantage as a novel detoxification mechanism.

59 BASIC BIOLOGICAL SCIENCES↗

Serum Neutralizing and Enhancing Effects on African Swine Fever Virus Infectivity in Adherent Pig PBMC

African swine fever virus (ASFV) causes hemorrhagic fever with mortality rates of up to 100% in domestic pigs. Currently, there are no commercial vaccines for the disease. Only some live-attenuated viruses have been able to protect pigs from ASFV infection. The immune mechanisms involved in the protection are unclear. Immune sera can neutralize ASFV but incompletely. The mechanisms involved are not fully understood. Currently, there is no standardized protocol for ASFV neutralization assays. In this study, a flow cytometry-based ASFV neutralization assay was developed and tested in pig adherent PBMC using a virulent ASFV containing a fluorescent protein gene as a substrate for neutralization. As with previous studies, the percentage of infected macrophages was approximately five time higher than that of infected monocytes, and nearly all infected cells displayed no staining with anti-CD16 antibodies. Sera from naïve pigs and pigs immunized with a live-attenuated ASFV and fully protected against parental virus were used in the assay. The sera displayed incomplete neutralization with MOI-dependent neutralizing efficacies. Extracellular, but not intracellular, virions suspended in naïve serum were more infectious than those in the culture medium, as reported for some enveloped viruses, suggesting a novel mechanism of ASFV infection in macrophages. Both the intracellular and extracellular virions could not be completely neutralized.

59 BASIC BIOLOGICAL SCIENCES↗

Dissecting Phenotype from Genotype with Clinical Isolates of SARS-CoV-2 First Wave Variants

The emergence and availability of closely related clinical isolates of SARS-CoV-2 offers a unique opportunity to identify novel nonsynonymous mutations that may impact phenotype. Global sequencing efforts show that SARS-CoV-2 variants have emerged and then been replaced since the beginning of the pandemic, yet we have limited information regarding the breadth of variant-specific host responses. Using primary cell cultures and the K18-hACE2 mouse, we investigated the replication, innate immune response, and pathology of closely related, clinical variants circulating during the first wave of the pandemic. Mathematical modeling of the lung viral replication of four clinical isolates showed a dichotomy between two B.1. isolates with significantly faster and slower infected cell clearance rates, respectively. While isolates induced several common immune host responses to infection, one B.1 isolate was unique in the promotion of eosinophil-associated proteins IL-5 and CCL11. Moreover, its mortality rate was significantly slower. Lung microscopic histopathology suggested further phenotypic divergence among the five isolates showing three distinct sets of phenotypes: (i) consolidation, alveolar hemorrhage, and inflammation, (ii) interstitial inflammation/septal thickening and peribronchiolar/perivascular lymphoid cells, and (iii) consolidation, alveolar involvement, and endothelial hypertrophy/margination. Together these findings show divergence in the phenotypic outcomes of these clinical isolates and reveal the potential importance of nonsynonymous mutations in nsp2 and ORF8.

60 APPLIED LIFE SCIENCES↗

Rapid Detection and Quick Characterization of African Swine Fever Virus Using the VolTRAX Automated Library Preparation Platform

African swine fever virus (ASFV) is the causative agent of a severe and highly contagious viral disease affecting domestic and wild swine. The current ASFV pandemic strain has a high mortality rate, severely impacting pig production and, for countries suffering outbreaks, preventing the export of their pig products for international trade. Early detection and diagnosis of ASFV is necessary to control new outbreaks before the disease spreads rapidly. One of the rate-limiting steps to identify ASFV by next-generation sequencing platforms is library preparation. Here, we investigated the capability of the Oxford Nanopore Technologies’ VolTRAX platform for automated DNA library preparation with downstream sequencing on Nanopore sequencing platforms as a proof-of-concept study to rapidly identify the strain of ASFV. Within minutes, DNA libraries prepared using VolTRAX generated near-full genome sequences of ASFV. Thus, our data highlight the use of the VolTRAX as a platform for automated library preparation, coupled with sequencing on the MinION Mk1C for field sequencing or GridION within a laboratory setting. These results suggest a proof-of-concept study that VolTRAX is an effective tool for library preparation that can be used for the rapid and real-time detection of ASFV.

60 APPLIED LIFE SCIENCES↗

Comparison of Routes of Administration, Frequency, and Duration of Favipiravir Treatment in Mouse and Guinea Pig Models of Ebola Virus Disease

Favipiravir is a ribonucleoside analogue that has been explored as a therapeutic for the treatment of Ebola Virus Disease (EVD). Promising data from rodent models has informed nonhuman primate trials, as well as evaluation in patients during the 2013–2016 West African EVD outbreak of favipiravir treatment. However, mixed results from these studies hindered regulatory approval of favipiravir for the indication of EVD. This study examined the influence of route of administration, duration of treatment, and treatment schedule of favipiravir in immune competent mouse and guinea pig models using rodent-adapted Zaire ebolavirus (EBOV). A dose of 300 mg/kg/day of favipiravir with an 8-day treatment was found to be fully effective at preventing lethal EVD-like disease in BALB/c mice regardless of route of administration (oral, intraperitoneal, or subcutaneous) or whether it was provided as a once-daily dose or a twice-daily split dose. Preclinical data generated in guinea pigs demonstrates that an 8-day treatment of 300 mg/kg/day of favipiravir reduces mortality following EBOV challenge regardless of route of treatment or duration of treatments for 8, 11, or 15 days. This work supports the future translational development of favipiravir as an EVD therapeutic.

60 APPLIED LIFE SCIENCES↗

Impact of Selenium and Vitamin E Deficiency on Zika Virus Pathogenesis and Immune Response in Mice

Micronutrient status is recognized to influence host susceptibility to viral infections, yet its impact on Zika virus (ZIKV) pathogenesis remains incompletely understood. We investigated the effects of dietary selenium and combined selenium plus vitamin E deficiency on ZIKV infection outcomes in a type I interferon α/β receptor knockout (Ifnar1 −/− ) murine model. Mice maintained on deficient diets exhibited significantly lower neutralizing antibody titers and reduced levels of key antiviral cytokines (IFN-γ, TNF-α, IFN-α, IFN-β, IL-12p70, CCL5) compared to controls. Correspondingly, higher viral RNA loads were detected in the brains of double-deficient mice, which also experienced greater weight loss and increased mortality. Deep sequencing revealed no major differences in overall viral genome diversity across diet groups; however, specific mutations, including V330L and D67E in the E gene, and V360I in the NS3 gene, were enriched or detected in nutritionally deficient animals. These findings suggest that antioxidant micronutrient deficiency impairs both humoral and cellular immune responses to ZIKV, potentially facilitating enhanced neuroinvasion. While the functional consequences of the identified mutations warrant further investigation, our results underscore the importance of adequate micronutrient intake for optimal antiviral defense. Further studies are needed to clarify the epidemiological significance of these observations.

Biological and medical sciences↗

Protection against Different Genotypes of Newcastle Disease Viruses (NDV) Afforded by an Adenovirus-Vectored Fusion Protein and Live NDV Vaccines in Chickens

The efficacy of an adenovirus-vectored Newcastle disease virus (NDV) vaccine expressing the fusion (F) NDV protein (adeno-F) was evaluated against challenges with virulent heterologous and homologous NDV strains to the F protein. In a preliminary study, two different doses (low and high) of adeno-F were tested against a virulent NDV strain containing the homologous NDV F protein, CA02. In a second study, at three weeks post-vaccination, the efficacy of the high dose of adeno-F was compared to a live attenuated NDV vaccine strain (LaSota) against three antigenically distinct virulent NDV challenge strains, one homologous (CA02) and two heterologous (TZ12, EG14) to F in the vectored vaccine. In both experiments, clinical signs, mortality, virus shedding, and humoral response were evaluated. In the first experiment, the survival rates from birds vaccinated with adeno-F at a high and low dose were 100% and 25%, respectively. In the second experiment, birds vaccinated with the high dose of adeno-F had a survival rate of 80%, 75%, and 65% after challenge with the CA02, TZ12, and EG14 viruses, respectively. All of the LaSota-vaccinated birds survived post-challenge no matter the NDV challenge strain. High antibody titers were detected after vaccination with LaSota by HI and ELISA tests. The majority of adeno-F-vaccinated birds had detectable antibodies using the ELISA test, but not using the HI test, before the challenge. The data show that both the similarity of the F protein of the adeno-F vaccine to the challenge virus and the adeno-F vaccination dose affect the efficacy of an adenovirus-vectored NDV vaccine against a virulent NDV challenge.

Ferreira, Helena L. (ORCID:0000000232613712)↗

Novel Protocol for Acute In Situ Ecotoxicity Test Using Native Crustaceans Applied to Groundwater Ecosystems

Current standardized laboratory test protocols use model species that have limitations to accurately assess native species responses to stressors. We developed and tested a novel acute in situ protocol for testing field-collected organisms. We used Asellus aquaticus and NaCl as a reference toxicant to test for the effects of location (laboratory vs. in situ), medium (synthetic vs. field water), substrate (presence vs. absence), and protocol replicability. We further tested the protocol using groundwater-adapted isopods: Proasellus assaforensis for the effect of location, P. cavaticus of medium and P.lusitanicus of substrate. Our results showed that A.aquaticus’ lethality obtained with the novel acute in situ protocol did not significantly differ from those from laboratory testing. However, laboratory tested P.assaforensis showed a higher sensitivity, suggesting that its acclimation to laboratory conditions might have pernicious effects. A. aquaticus and P. cavaticus showed a higher mortality using synthetic medium in situ and under laboratory conditions, which overestimated the stressor’s effect. Besides, substrate use had no significant effect. The novel acute in situ protocol allows the use of native species under realistic scenarios. It is particularly well adapted for assessing the risk of groundwater ecosystems but it can be applied to a wide range of ecosystems.

Castaño-Sánchez, Andrea↗

Examination of an Electrified Bar Rack Fish Guidance Device for Hydropower Turbines

The potential of hydropower turbines to cause injury or mortality to fish is a concern. To reduce this risk, engineers have begun to develop a conically arranged, cantilevered electrified bar rack (Center Sender). This device is proposed to be mounted within the intake of a turbine, guiding downstream-passing fish towards the center of the turbine where blade velocities are lower and blades are thicker, likely reducing the potential for fish to be injured. A simplified version was installed in a flume for testing with Rainbow trout (Oncorhynchus mykiss) while examining several parameters such as electrification, bar spacing, bar angle, and water velocity. The most effective settings were observed to be a water velocity of 1.0 m s -1 with all bars installed at an angle of 40° with electrification on. Other combinations were still effective but had varying results. A minimal proportion (2.3%) of fish passed at lower velocities with bars electrified and mounted at 20°, suggesting that while it functions well as a guidance device in high-velocity conditions, it performs better as a barrier at lower velocities. The study suggests that the Center Sender has promising potential for reducing the harmful effects of hydropower turbines on fish populations by guiding them away from stressors. Further research is needed, but current results encourage its potential use in hydropower operations.

13 HYDRO ENERGY↗

The Effect of Size on Postrelease Survival of Head-Started Mojave Desert Tortoises

Abstract Captive-rearing conservation programs focus primarily on maximizing postrelease survival. Survival increases with size in a variety of taxa, often leading to the use of enhanced size as a means to minimize postrelease losses. Head-starting is a specific captive-rearing approach used to accelerate growth in captivity prior to release in the wild. We explored the effect of size at release, among other potential factors, on postrelease survival in head-started Mojave desert tortoises Gopherus agassizii. Juvenile tortoises were reared for different durations of captivity (2–7 y) and under varying husbandry protocols, resulting in a wide range of juvenile sizes (68–145 mm midline carapace length) at release. We released all animals (n = 78) in the Mojave National Preserve, California, United States, on 25 September 2018. Release size and surface activity were the only significant predictors of fate during the first year postrelease. Larger sized head-starts had higher predicted survival rates when compared with smaller individuals. This trend was also observed in animals of the same age but reared under different protocols, suggesting that accelerating the growth of head-started tortoises may increase efficiency of head-starting programs without decreasing postrelease success. Excluding five missing animals, released head-starts had 82.2% survival in their first year postrelease (September 2018–September 2019), with all mortalities resulting from predation. No animals with >90-mm midline carapace length were predated by ravens. Our findings suggest the utility of head-starting may be substantially improved by incorporating indoor rearing to accelerate growth. Target release size for head-started chelonians will vary among head-start programs based on release site conditions and project-specific constraints.

Biodiversity & Conservation↗

Design of Broadly Cross-Reactive M Protein–Based Group A Streptococcal Vaccines

Group A streptococcal infections are a significant cause of global morbidity and mortality. A leading vaccine candidate is the surface M protein, a major virulence determinant and protective Ag. An obstacle to the development of M protein–based vaccines is the >200 different M types defined by the N-terminal sequences that contain protective epitopes. Despite sequence variability, M proteins share coiled-coil structural motifs that bind host proteins required for virulence. In this study, we exploit this potential Achilles heel of conserved structure to predict cross-reactive M peptides that could serve as broadly protective vaccine Ags. Combining sequences with structural predictions, six heterologous M peptides in a sequence-related cluster were predicted to elicit cross-reactive Abs with the remaining five nonvaccine M types in the cluster. The six-valent vaccine elicited Abs in rabbits that reacted with all 11 M peptides in the cluster and functional opsonic Abs against vaccine and nonvaccine M types in the cluster. We next immunized mice with four sequence-unrelated M peptides predicted to contain different coiled-coil propensities and tested the antisera for cross-reactivity against 41 heterologous M peptides. Based on these results, we developed an improved algorithm to select cross-reactive peptide pairs using additional parameters of coiled-coil length and propensity. The revised algorithm accurately predicted cross-reactive Ab binding, improving the Matthews correlation coefficient from 0.42 to 0.74. These results form the basis for selecting the minimum number of N-terminal M peptides to include in potentially broadly efficacious multivalent vaccines that could impact the overall global burden of group A streptococcal diseases.

60 APPLIED LIFE SCIENCES↗

Assessing the representation of the Australian carbon cycle in global vegetation models

Australia plays an important role in the global terrestrial carbon cycle on inter-annual timescales. While the Australian continent is included in global assessments of the carbon cycle such as the global carbon budget, the performance of dynamic global vegetation models (DGVMs) over Australia has rarely been evaluated. We assessed simulations of net biome production (NBP) and the carbon stored in vegetation between 1901 to 2018 from 13 DGVMs (TRENDY v8 ensemble). We focused our analysis on Australia's short-term (inter-annual) and long-term (decadal to centennial) terrestrial carbon dynamics. The TRENDY models simulated differing magnitudes of NBP on inter-annual timescales, and these differences resulted in significant differences in long-term vegetation carbon accumulation (–4.7 to 9.5 Pg C). We compared the TRENDY ensemble to several satellite-derived datasets and showed that the spread in the models' simulated carbon storage resulted from varying changes in carbon residence time rather than differences in net carbon uptake. Differences in simulated long-term accumulated NBP between models were mostly due to model responses to land-use change. The DGVMs also simulated different sensitivities to atmospheric carbon dioxide (CO 2 ) concentration, although notably, the models with nutrient cycles did not simulate the smallest NBP response to CO 2 . Our results suggest that a change in the climate forcing did not have a large impact on the carbon cycle on long timescales. However, the inter-annual variability in precipitation drives the year-to-year variability in NBP. We analysed the impact of key modes of climate variability, including the El Niño–Southern Oscillation (ENSO) and the Indian Ocean Dipole (IOD), on NBP. While the DGVMs agreed on sign of the response of NBP to El Niño and La Niña and to positive and negative IOD events, the magnitude of inter-annual variability in NBP differed strongly between models. In addition, we find that differences in the timing of simulated phenology and fire dynamics are associated with differences in simulated or prescribed vegetation cover and process representation. We further find model disagreement in simulated vegetation carbon, phenology, and apparent carbon residence time, indicating that the models have different types and coverage of vegetation across Australia (whether prescribed or emergent). Our study highlights the need to evaluate parameter assumptions and the key processes that drive vegetation dynamics, such as phenology, mortality, and fire, in an Australian context to reduce uncertainty across models.

54 ENVIRONMENTAL SCIENCES↗

Exploring the impacts of unprecedented climate extremes on forest ecosystems: hypotheses to guide modeling and experimental studies

Abstract. Climatic extreme events are expected to occur more frequently in the future, increasing the likelihood of unprecedented climate extremes (UCEs) or record-breaking events. UCEs, such as extreme heatwaves and droughts, substantially affect ecosystem stability and carbon cycling by increasing plant mortality and delaying ecosystem recovery. Quantitative knowledge of such effects is limited due to the paucity of experiments focusing on extreme climatic events beyond the range of historical experience. Here, we present a road map of how dynamic vegetation demographic models (VDMs) can be used to investigate hypotheses surrounding ecosystem responses to one type of UCE: unprecedented droughts. As a result of nonlinear ecosystem responses to UCEs that are qualitatively different from responses to milder extremes, we consider both biomass loss and recovery rates over time by reporting a time-integrated carbon loss as a result of UCE, relative to the absence of drought. Additionally, we explore how unprecedented droughts in combination with increasing atmospheric CO2 and/or temperature may affect ecosystem stability and carbon cycling. We explored these questions using simulations of pre-drought and post-drought conditions at well-studied forest sites using well-tested models (ED2 and LPJ-GUESS). The severity and patterns of biomass losses differed substantially between models. For example, biomass loss could be sensitive to either drought duration or drought intensity depending on the model approach. This is due to the models having different, but also plausible, representations of processes and interactions, highlighting the complicated variability of UCE impacts that still need to be narrowed down in models. Elevated atmospheric CO2 concentrations (eCO2) alone did not completely buffer the ecosystems from carbon losses during UCEs in the majority of our simulations. Our findings highlight the consequences of differences in process formulations and uncertainties in models, most notably related to availability in plant carbohydrate storage and the diversity of plant hydraulic schemes, in projecting potential ecosystem responses to UCEs. We provide a summary of the current state and role of many model processes that give way to different underlying hypotheses of plant responses to UCEs, reflecting knowledge gaps which in future studies could be tested with targeted field experiments and an iterative modeling–experimental conceptual framework.

54 ENVIRONMENTAL SCIENCES↗