Abstract: The cactus pear (Opuntia ficus-indica) is a crucial plant in Tigray, northern Ethiopia, widely distributed in arid and semi-arid environments. It serves as a seasonal food, livestock feed, fence, soil conservation, and environmental protection. Recently, the cactus pear population in Tigray have been damaged by an exotic insect, cochineal (Dactylopius coccus). It damaged the cactus pear populations in the region’s southern, south eastern and eastern zones. The Tigray war that broke out in November 2021 exacerbated D. coccus infestation. A study was conducted to assess the impact of the armed conflict on the trends of D. coccus infestation and forward management approaches for sustainable cactus pear production in post-war Tigray. The study was conducted in the eastern zone of Tigray. Primary and secondary data were collected. The study revealed that D. coccus infestation increased during and post-war compared to the pre-war. The number of districts and level of D. coccus¬ infestation on cactus pear populations increased. The rapid D. coccus spread is attributed to the impact of the armed conflict which halted the pest management practices. Different management approaches are recommended to manage D. coccus dissemination and sustainably produce cactus pear in the region, including pest prevention, suppression, or eradication.

Abstract: Due to adsorption of all metal ions (and some non-metals like As, Sb), metal complexes and metal-rich small (oxide, carbonate, oxalate) particles alike, chitin does remove these M spe-cies from a solution or from moist sediments, depleting the solution down to pMol/l levels except for alkali metals, or Mg. Chitin retains substantial metal ions from solutions < 1 nMol/l. Accordingly, there is a (concentration gradient-caused) voltage between two identi-cal pure metal or -alloy electrodes of which one is wrapped by chitin. While 10 min will do to obtain reproducible adsorption on chitin, electrochemical measurements show that the voltage forming between a chitin-surrounded and an identical bare metal electrode keeps on changing for 24 hours except for Ni. Preliminary experiments showed that dried arthropods produce similar results as previously purified chitin(proteins and metal carbonates removed during workup). On chitin exist on-equivalent binding sites the former of which are quickly populated and the metal ions then pass to the latter. However, these latter sites are still lo-cated next to the surface of chitin: upon addition of an appropriate ligand, voltage will mas-sively increase within 1 – 2 min which shows that even sizable ligand ions (glycinate, phe-nolic carboxylic acids like caffeic acid) can still access the metal ions in aq. solution. Chitin does catalyze ligand glycinate oxidation by introduced air oxygen next to cobalt electrodes. Multimetal electrode chitin-modified sensor systems, batteries and designs of fuel cells are discussed. This does agree with results concerning adsorption on surface of living crayfish and crickets and means that the latter can be involved in investigation protocols for environmental pollu-tants. Except for a longer lag time, electrochemical data are easier to obtain than analyzing metal contents on a chitin interface. Voltage does increase rapidly when there is addition of either a ligand, some species connecting a metal ion to the chitin surface by some molecular or ionic bridge or of an anion which does form hardly soluble salts (e.g., to set M levels in solution similar to those in the open environment) and keeps up for several days until the chitin gets saturated with the metal ion. Oxidation of ligand (glycinate) or reduction of pre-cipitating agent (iodate) both change the voltage but with different signs. Effects of SCN- addition suggest possible photoactivation of signals, too.

Abstract: Melanoma is the most common type of skin cancer. Melanomas are well-known for their ability to metastasize to other organs, including the lungs, liver, brain, and bones. The ability of melanoma cells to switch among different phenotypes is a key mechanism that underscores their metastatic potential. We report here on the effect of calcium sulfide (CaS) dispersions in melanoma cells. Melanomas with the mesenchymal and epithelial phenotypes were observed during cell culture preparation. The dispersion reduced the relative percentage of melanomas with the mesenchymal and epithelial phenotypes to (55 + 5) and (57 + 5) %, respectively. In contrast, the viability of normal fibroblasts treated with the dispersion, or the reactants used to prepare the dispersion, remains nearly constant in the same period. Fluorescence imaging measurements are consistent with the release of cytochrome c from mitochondria and its translocation to the cell nuclei. The average expression of caspases 3 and 9 were found to be higher than in the corresponding melanoma control, consistent with intrinsic apoptosis. The response of vinculin expression was slightly different in types of cells. Vinculin was found to delocalize in the cytoplasm of treated mesenchymal melanoma cells with a slightly higher concentration at the end of the actin fibers. A statistically significant increase in the number of FAP in the edge of the cell membrane-external cellular matrix (ECM) was observed in post treated melanoma that exhibited the epithelial phenotype. The changes in vinculin expression and in the number of FAP are consistent with regulation of proteins associated with programmed cell death. It is proposed that the sulfides produced from the reactions of the nanoclusters in the acidic environment facilitate the regulation of proteins required to initiate apoptosis, although other processes may also be involved. Extension of the efforts to animal models will be an important step to the commercialization of the dispersion for melanoma treatment.

Abstract: This literature review examines the function and evolution of cranial crests in Dilophosaurus wetherilli, a notable theropod dinosaur from the Early Jurassic period. The review synthesizes morphological data and behavioral theories to provide a comprehensive analysis of the crests' roles in the dinosaur's life. Cranial crests, characterized by their distinctive, elongated structures extending from the skull, have intrigued paleontologists due to their unique appearance and apparent function. This review utilizes findings from recent fossil discoveries, comparative anatomy, and biomechanical studies to explore hypotheses regarding the crests' primary functions. Key theories suggest these structures may have served for display purposes, thermoregulation, or defensive mechanisms. Evidence from cranial and postcranial anatomy, as well as comparisons with extant analogs, provides insights into the possible evolutionary pressures influencing the development of crests in Dilophosaurus wetherilli. Additionally, the review considers the role of sexual dimorphism and social behavior in shaping these features. By integrating these diverse lines of evidence, the review aims to offer a nuanced understanding of how cranial crests might have contributed to the survival and social interactions of Dilophosaurus. This analysis not only advances our knowledge of Dilophosaurus wetherilli but also contributes to broader discussions on the evolution and functional morphology of early theropods.

Abstract: Human growth hormone (GH) exerts its pleiotropic effects by binding to its receptor (GHR), leading to receptor dimerization and activation. We combined structural, evolutionary, and genetic analyses to elucidate the critical determinants of GH-GHR interaction and the impact of disease-causing mutations. Protein contact analysis revealed the specific amino acid residues involved in two distinct binding interfaces between GH and two chains of GHR. ConSurf analysis demonstrated significant sequence conservation in the receptor-binding regions of GH across species, highlighting their functional importance. A comprehensive list of known disease-causing mutations in GH was compiled and mapped to these binding interfaces and conserved regions. Computational site-directed mutagenesis (SDM) analysis predicted the impact of several mutations on protein stability, revealing both stabilizing and destabilizing effects. Sequence comparisons with orthologs from various species further supported the evolutionary conservation of key functional residues. Integrated analysis of contact residues between GH and GHR showed a strong correlation between receptor-binding residues, evolutionary conservation, and the occurrence of disease-associated mutations. These findings underscore the critical role of specific GH residues in mediating high-affinity interaction with its receptor, and how mutations in these conserved contact points can disrupt binding affinity and/or protein stability, ultimately leading to growth disorders. This multi-faceted approach provides valuable insights into the molecular mechanisms underlying growth hormone deficiency and related syndromes.

Abstract: The homotrimeric P2X7 receptor (P2X7R) contains three ATP4- binding sites in its ectodomain. Here, we investigated the role of the individual ATP4- activation sites of the rat P2X7R (rP2X7R) using trimeric rP2X7R concatamers consisting either of three wild-type subunits (7-7-7) or concatamers with up to three subunits having knocked-out ATP binding sites (7ko-7ko-7ko). Following expression in Xenopus oocytes, ATP4--elicited ion currents were recorded using the two-microelectrode voltage clamp technique. The 7-7-7 concatamer exhibited a biphasic ATP4- concentration dependence, best fit by the sum of two Hill functions, confirming the existence of functionally distinct ATP4- activation sites. The activation time course of the 7-7-7 was best approximated by the sum of a fast and a slow exponential saturating activation component. Similarly, deactivation exhibited both a fast and a slow exponential decay. Only one Hill function was required to best fit the ATP4- concentration dependence of concatamers with only two or one ATP4- binding sites, and their deactivation time courses largely lacked the slowly deactivating components. We conclude that binding of one ATP4- is sufficient for partial activation of the rP2X7R and that allosteric effects occur when all three ATP4- binding sites are occupied, leading to distinct functional activation sites.

Abstract: This study examines the use of analgesic medications among sheep farmers, highlighting both their benefits and significant gaps in their adoption. Our findings indicate that 26.7% of sheep farmers did not utilize any analgesics, a figure that contrasts sharply with high usage rates exceeding among farmers in swine and cattle sectors. Low utilization of NSAIDs and corticosteroids in sheep farming may be attributed to inadequate education and the absence of approved indications for these medications in Greece. Additionally, we found a positive correlation between farm size and the use of NSAIDs, with larger farms using more NSAIDs, especially flunixin. Furthermore, farms using more NSAIDs/larger farms tended to have higher milk yields, suggesting that enhanced management practices are associated with better outcomes. The results underscore the need for targeted educa-tional initiatives to improve the use of analgesics in sheep farming and address welfare concerns. Overall, this research highlights the importance of addressing gaps in pain management practices within the sheep farming sector to promote better animal welfare. However, further studies with larger sample sizes are recommended to expand our un-derstanding of analgesic use and its implications for sheep welfare.

Abstract: Simian foamy virus (SFV) is a retrovirus that infects primates, including American primates (AP), but epidemiological studies are often limited to captive animals. The state of Rio de Janeiro (RJ) is home to an endemic AP species, Leontopithecus rosalia, and an invasive species, Leontopithecus chrysomelas, both endangered. This study assessed the molecular prevalence of SFV in these species. Genomic DNA was extracted from 48 oral swab samples of L. chrysomelas (Niterói/RJ) and 102 of L. rosalia (Silva Jardim/RJ). qPCR was performed to diagnose and evaluate proviral load (pVL). SFV prevalence was 23% in L. chrysomelas and 33% in L. rosalia. No age-related differences were observed, but L. rosalia showed a higher average pVL (3.27 log10/10⁶ cells) compared to L. chrysomelas (3.03 log10/10⁶ cells) (p=0.005). The viral sequence of L. rosalia clustered within a monophyletic SFVlro clade, distinct from two SFVlchrysom lineages. Origin of SFVlro dates back to 79.6 thousands of years ago. This was the first study to determine the molecular prevalence of SFV in free-living populations of Leontopithecus, and may be of great importance for elucidating the complex evolutionary history of SFV in AP.

Abstract: Background/Objectives: Parietin (PTN), a blue-light absorbing pigment from Teloschistes spp. lichens, exhibits photosensitizing properties via Type I (superoxide anion, O2•-) and Type II (singlet oxygen, 1O2) mechanism, inactivating bacteria in vitro after photo-stimulation. We evaluate the in vitro antifungal activity of PTN against Candida tropicalis biofilms under actinic irradiation, its role in O2•- and 1O2 production, and the cellular stress response. Methods: Minimum Inhibitory Concentration (MIC) of PTN was determined in C. tropicalis NCPF 3111 under dark and actinic light conditions. Biofilm susceptibility was assessed at MIC/2, MIC, MICx2, MICx4 and MICx6 in the same conditions, and viability was measured by colony forming units. Photodynamic mechanisms were examined using Tiron (O2•- quencher) or sodium azide (1O2 scavenger). O2•- production was measured by the Nitro-Blue tetrazolium (NBT) reduction and nitric oxide (NO) by Griess assay. Total antioxidant capacity was studied by FRAP (Ferrous Reduction Antioxidant Potency) assay and superoxide dismutase (SOD) activity by NBT assay. Results: Photo-stimulation of PTN reduced C. tropicalis biofilm viability by 4-logs at MICx2. Sodium azide partially reversed the effect, whereas Tiron fully inhibited it, indicating the critical role of O₂•⁻. PTN also increased O₂•⁻ and NO levels, enhancing SOD activity and FRAP. However, this antioxidant response was insufficient to prevent biofilm photoinactivation. Conclusions: Photoinactivation of C. tropicalis biofilms by PTN is primarily mediated by O2•-, with a minor contribution from ¹O₂ and an imbalance in NO levels. These findings suggest PTN as a promising photosensitizer for antifungal photodynamic therapy.

Abstract: System suitability testing of chromatography is an indispensable procedure in pharmaceutical analysis, and must comply with rules in related pharmacopoeias. An inverse Fourier transform algorithm is developed to accurately evaluate chromatographic features versus a standard Gaussian peak shape. The regular chromatogram is considered as a pseudo-frequency spectrum and can be converted to a nominal time signal by inverse Fourier transformation. The system suitability parameters of peak width, theoretical plate number, tailing factor and noise testing were evaluated by linear regressions directly and compared with the compendial rules. This novel method is simple, accurate, robust, reliable and efficient for evaluation of chromatographic peak features.

Abstract: A novel temporally resolved settle-plate air sampler was developed using 3D printing technology to improve upon traditional passive air sampling methods. Conventional settle plates provide cumulative measurements of particle or microbial loads over an entire sampling period, lacking temporal resolution necessary to identify specific contamination events. The described device integrates a Petri plate within a 3D-printed housing featuring a narrow slit that exposes only a small portion of the plate to incoming particles. A rotary mechanism, driven by a mechanical clock motor, rotates the petri plate over 12 hours, allowing for time-segmented sampling. Validation experiments demonstrated the device's ability to accurately encode the temporal history of particle deposition using both aerosolized dyes and viable microbial spores. The device effectively correlated bioaerosol deposition with ambient wind conditions during outdoor sampling. The system is inexpensive (under $10 USD), requires no specialized skills to assemble, and is compatible with existing settle plate methodologies. This innovation enhances the ability to conduct air quality assessments in critical environments, enabling data-driven decisions to mitigate contamination risks.

Abstract: Accurate, high-throughput canopy phenotyping using UAV-based multispectral remote sensing is critically important for optimizing the management and breeding of foxtail millet in rainfed regions. This study integrated multi-temporal field measurements of leaf water content, SPAD-derived chlorophyll, and leaf area index (LAI) with UAV imagery (red, green, red-edge, near-infrared bands) across two sites and two consecutive years (2023 and 2024) in Shanxi Province, China. Various modeling approaches, including Random Forest, Gradient Boosting, and regularized regressions (e.g., Ridge, Lasso), were evaluated for cross-regional and cross-year extrapolation. Results showed that single-site modeling achieved coefficients of determination (R²) up to 0.95, with mean relative errors of 10%–15% in independent validations. When models were transferred between sites, R² generally remained between 0.50 and 0.70, although SPAD estimates exhibited larger deviations under high-nitrogen conditions. Even under severe drought in 2024, cross-year predictions still attained R² values near 0.60. Among these methods, tree-based models demonstrated strong capability for capturing nonlinear canopy trait dynamics, whereas regularized regressions offered simplicity and interpretability. Incorporating multi-site and multi-year data further enhanced model robustness, increasing R² above 0.80 and markedly reducing average prediction errors. These findings demonstrate that rigorous radiometric calibration and appropriate vegetation index selection enable reliable UAV-based phenotyping for foxtail millet under diverse environments and time frames. The proposed approach thus provides strong technical support for precision management and cultivar selection in semi-arid foxtail millet production systems.

Abstract: Corynebacterium pseudotuberculosis is a non-motile, β-hemolytic bacterium and causative factor of caseous lymphadenitis. The disease affects sheep and goats causing impaired wool production, weight loss and carcass condemnation. Our previous work has elucidated the morphology, heterogeneity, and antimicrobial susceptibility of C. pseudotuberculosis biofilm. However, the information of proteome expression underlying C. pseudotuberculosis biofilm development remains scarce. Thus, the objective of the present work is to compare the whole-cell proteome profiles between planktonic and biofilm fractions of C. pseudotuberculosis and identify C. pseudotuberculosis proteins and biological pathways showing differential expressions. C. pseudotuberculosis biofilm was grown in a six-well microplate for 24 h at 37°C. Polyacrylamide gel electrophoresis combined with tandem mass spectrometry and bioinformatics analysis was conducted to analyze proteome expression. Results demonstrated differential expression of seven SDS-PAGE protein bands (33.7 – 150 kDa) in comparison between the planktonic and biofilm fractions of C. pseudotuberculosis. Overall, 711 proteins that showed differential expression were successfully identified while the protein-protein interaction network revealed a total of 1206 functional linkages among the differentially expressed proteins. Fifty-seven hub proteins with more than 10 functional linkages were identified, including large subunit ribosomal protein L3, translation initiation factor IF-2, multifunctional oxoglutarate decarboxylase and DNA-dependent RNA polymerase. Functional enrichment analysis showed the association of differentially expressed C. pseudotuberculosis proteins with amino acid metabolism (p-value: 0.0243) and secondary metabolite metabolism (p-value: 0.0198). In conclusion, the formation of C. pseudotuberculosis biofilm may involve changes in multiple metabolic pathways.

Abstract: Weed suppression is a crucial factor in sustainable agriculture, and optimizing plant mixtures can enhance weed control efficiency. This study evaluates the effects of different mixture ratios of common vetch (Vicia sativa L.) and annual ryegrass (Lolium multiflorum L.) on forage yield, biomass production dry matter production, and weed suppression in organic forage cropping systems. Field experiments were conducted during the 2021-2022 growing season at two locations in Turkey: Ankara/Yenikent and Manisa/Beydere, using 11 mixture ratios ranging from 100% vetch to 100% ryegrass. Results showed that ryegrass-dominant mixtures, particularly 10% vetch / 90% ryegrass and 30% vetch / 70% ryegrass, achieved the highest forage and dry matter yields while maintaining effective weed suppression. Pure ryegrass systems (100% ryegrass) exhibited the highest overall productivity, whereas pure vetch (100% vetch) treatments were less effective in weed control and biomass production. Environmental differences between locations significantly influenced the performance of mixtures, with Manisa/Beydere yielding higher overall results. This study highlights the potential of optimizing vetch-ryegrass mixtures to balance forage yield, weed suppression, and adaptability in organic cropping systems, offering practical insights for sustainable forage production.

Abstract: Porcine Epidemic Diarrhea Virus (PEDV) was introduced in the United States (U.S.) in 2013, spreading rapidly and leading to economic losses. Two strains, S-INDEL and non-S-INDEL, are present in the U.S. We analyzed 313 genomes and 556 Spike protein sequences generated since its introduction. PEDV case numbers were highest during the first two years after its introduction (epidemic phase), then declined and stabilized in the following years (endemic phase). Sequence availability was higher during the initial epidemic phase. Our results suggest non-S-INDEL strain is the predominant strain in U.S. The non-S-INDEL strain exhibits a nucleotide identity percentage above 97.6%. Most non-S-INDEL sequences sampled after 2017 clustered into two sub-clades. No descendants derived from other clades present in the epidemic period were identified in the contemporary data, suggesting that these clades are no longer circulating in the U.S. The two clades currently circulating are restricted to two respective geographic regions and our results suggest limited inter-regional spread. This insight helps determine the risk of re-introduction of PEDV if it were regionally eliminated. Continued molecular surveillance is key to confirming the extinction of older clades, mapping the distribution and spread of recent clades, and understanding PEDV's evolutionary diversification..

Abstract: Pine wilt disease, caused by the nematode Bursaphelenchus xylophilus, poses a significant threat to pine forests worldwide. Understanding the dynamics of its spread is crucial for effective disease management. In this study, we investigated the involvement of asymptomatic carrier trees in the expansion of pine wilt disease through a series of experiments. Cage-releasing experiments revealed that sexually immature Japanese pine sawyer, Monochamus alternatus feeding on healthy pine branches drop only minimal number of nematodes (primary infection). However, sexually mature beetles, still harboring numerous nematodes, fly to weakened trees for breeding and extend their feeding activities to healthy pines around weakened trees, infecting them with nematodes and thus spreading the disease further. Inoculation experiments on field-planted black pine seedlings demonstrated that even a small number of nematodes can lead to a high occurrence of asymptomatic carrier trees. Our findings suggest that nematode infections transmitted by sexually mature Monochamus beetles significantly contribute to the expansion of pine wilt damage and play a crucial role in the persistence of asymptomatic carrier trees. This conclusion is based on cage release experiments demonstrating nematode transmission by mature beetles and inoculation experiments highlighting the conditions leading to asymptomatic carrier trees.

Abstract: Yarrowia lipolytica has a long history of industrial use. With its high secretion capacity, GRAS status and wide range of engineering tools, this yeast has become a powerful host for heterologous protein production as well as a favourite candidate for cellular factory design. Among integration vectors mostly used worldwide for Y. lipolytica engineering, zeta-based auto-cloning vectors took advantage of their bordering sequences (LTRs of Ylt1 retrotransposon) to integrate either by homology in Ylt1-bearing strains or non-homologously into any strain devoid of this retrotransposon. As such non-homologous integrations were less efficient and reliable, there was a need for designing zeta-bearing derivatives of widely used Y. lipolytica strains. Namely, the W29 wild-type strain and its genetically engineered derivative Po1f strain were equipped with a zeta docking platform in order to allow a more efficient and reliable integration of any zeta-based vector. The presence of this new docking platform allows to compare directly the performances of transformants obtained with different constructs, enabling to perform high-throughput screening or protein engineering. The efficiency of these new Po1z and Po1zP strains was demonstrated through the surexpression of two native proteins, a chitinase and a protease. At last, new zeta-based auto-cloning vectors with more powerful promoters were added to this new expression system.

Abstract: New psychoactive substances (NPS) pose a global public health challenge. Its ongoing evolution intensifies drug abuse cases, complicates regulation, and makes comprehensive studies difficult to conduct. Emerging studies indicate that these substances may severely impact spermatogenesis, reproductive hormones and male fertility. This review aims to consolidate existing knowledge on the impact of NPS on male reproduction, identify gaps in the literature, and suggest directions for future research. This comprehensive review examines original, peer-reviewed studies and clinical reports from PubMed, Semantic Scholar, AJOL, and Google Scholar. Data collection spanned from 2014 to 2024, using keywords such as “amphetamine-type stimulants”, “ATS”, “amphetamine”, “stimulants”, “opioids”, “opioid abuse”, “opioid addiction”, “male reproduction”, “male fertility”, and “reproductive toxicity”. The search encompasses illegal drugs, prescription medications, and over-the-counter (OTC) drugs that induce male reproduction toxicity through chronic use or prolonged exposure. Although current studies have limitations, our findings indicate that both illicit and medicinal ATS and opioids negatively impact male reproduction. These substances can diminish sperm quality, disrupt reproductive hormones, and cause sperm DNA damage. This review highlights further study in formulating public health strategies and supplements that mitigate DNA damage caused by these substances.

Abstract: (1) Background: This study aims to analyze the defoliation and boll opening performance of 123 upland cotton germplasm resources after spraying defoliant, using multispectral data to select relevant vegetation indices and identify germplasm re-sources sensitive to defoliants, providing methods for cotton variety improvement and high-quality parental resources. (2) Methods: 123 historical upland cotton germplasm resources from Xinjiang were selected, and the defoliation and boll opening of cotton leaves were investigated at 0, 4, 8, 12, 16, and 20 days after defoliant application. Sim-ultaneously, multispectral digital images were collected using drones to obtain 12 vegetation indices. Based on defoliation rate, the optimal vegetation index was selected, and defoliant-sensitive germplasm resources were identified. (3) Results: The most significant difference in defoliation rate of cotton germplasm resources occurred 16 days after application. Cluster analysis grouped the 123 breeding materials into three categories, with Class I showing the best defoliation effect. Among the 12 vegetation indices, the Plant Senescence Reflectance Index (PSRI) has the highest correlation coef-ficient with the defoliation rate; and when the PSRI value is higher, the defoliation ef-fect of the material is better. Using drone multispectral technology, 15 defoli-ant-sensitive cotton materials were identified, with defoliation rates exceeding 85%, boll opening rates ranging from 76.67% to 98.04%, and PSRI values between 0.1607 and 0.1984.(4) Conclusions: The study found that vegetation indices with sensitive re-sponses can serve as effective indicators for evaluating the sensitivity of cotton breed-ing materials to defoliants. The combined analysis of traditional survey methods and PSRI classification demonstrates that using drone multispectral technology as a sub-stitute for manual methods in large-scale, rapid monitoring and selection of cotton breeding materials with excellent defoliation effects is feasible.

Abstract: To ensure sustainability, the Ecosystem Approach to Fisheries (EAF) requires the evaluation of the impacts of fisheries beyond the main targeted species, to include those on bycaught, endangered threatened and protected populations and keystone species. However, traditional stock assessments require extensive datasets that are often unavailable for data-limited fisheries, particularly in small-scale settings or the Global South. This study evaluates the robustness of length-based approaches for fish stock assessment by comparing simple indicators and quantitative methods using an age-structured Operating Model. Simulations were conducted for a range of scenarios, for a range of life-history types and recruitment and natural mortality dynamics. Results reveal that while length-based approaches can effectively track trends in fishing mortality, performance varies significantly depending on species-specific life histories and assumptions about key parameters. Simple indicators often matched or outperformed complex methods, particularly when assumptions about equilibrium conditions or natural mortality were violated. The study highlights the limitations of length-based methods for classifying stock status relative to reference points, but demonstrates their utility when used with historical reference periods or as part of empirical harvest control rules. The findings provide practical guidance for applying length-based approaches in data-limited fisheries management, ensuring sustainability in data and capacity situations.