Cadmium (Cd) is a prevalent and toxic heavy metal widely present in soils. Cotton (Gossypium hirsutum L.), which readily accumulates Cd, exhibits inhibited seed germination under Cd stress. Melatonin (MT), a potent indoleamine antioxidant, has been demonstrated to facilitate seed germination under adverse conditions, but its regulatory mechanisms in cotton under Cd stress remain unclear. In this study, cotton seeds of the cultivar 'Nongda 601' were used to investigate the effects of exogenous MT on germination characteristics, antioxidant enzyme activities, and hormone levels under Cd stress. Experimental treatments included a control (CK), MT treatment, Cd treatment, and a combined Cd + MT treatment (CM). Transcriptome sequencing was further conducted to elucidate the underlying mechanisms. Cd stress significantly inhibited cotton seed germination, reduced amylase activity, and increased starch accumulation. It also decreased antioxidant enzyme activities while increasing reactive oxygen species (ROS) and malondialdehyde (MDA) contents. In addition, Cd stress increased IAA, ABA, and endogenous MT contents, while decreasing GA3 and ZR levels. Exogenous MT significantly enhanced amylase and antioxidant enzyme activities, decreased ROS, MDA, and starch contents, and modulated hormone levels by increasing IAA, GA3, ZR, and endogenous MT, while decreasing ABA content. Transcriptome analysis indicated that MT upregulated the expression of antioxidant-related genes under Cd stress, including SODA, FSD, CAT2, and pod. Additionally, exogenous MT alleviated Cd toxicity by regulating hormone signal transduction and the phenylpropanoid biosynthesis pathway. Exogenous MT alleviates Cd-induced inhibition of cotton seed germination by enhancing antioxidant capacity, promoting starch mobilization, and rebalancing hormone signaling, while modulating phenylpropanoid metabolism. These findings provide a theoretical foundation for improving cotton cultivation in Cd-contaminated soils.

Bio-coating is a process of coating seed surfaces with plant-beneficial microorganisms, allowing them to colonize plant roots from the very beginning of plant growth to protect the plant from diseases. Okra and cotton seeds were double-coated with biocontrol agents using different polymer combinations to suppress phytopathogens. The bio-coated seeds were evaluated for germination percentage and viability of bioinoculants during storage, and later for plant growth and disease suppression under pot house conditions. Okra coated with Trichoderma asperellum using a lab-prepared combination of chitosan and PVP (Polymer mix- T -Cts-PVP) (Cts: chitosan, PVP: polyvinyl pyrrolidone, T: Trichoderma asperellum) maximally inhibited the growth of Alternaria alternata (73.68%), Rhizoctonia solani (51.80%), and Fusarium oxysporum (48.42%). Similar observations were recorded for cotton seeds coated with Trichoderma asperellum and bacterial consortium (Azotobacter chroococcum HT-54, Bacillus subtilis, Pseudomonas fluorescens, Stenotophomonas maltophilia, and Bacillus megaterium) using carboxymethyl cellulose (CMC) and PVP. All bio-coated seeds showed better germination percentages than the control. All the polymers CMC, chitosan and PVP (polymer mix-T-CMC-PVP and polymer mix-T-Cts-PVP), were more compatible with Trichoderma asperellum than with the bacterial consortium and retained more viable spores for three months on okra seeds. While opposite was observed for cotton seeds and only polymer mix-B-CMC-PVP and polymer mix-T-CMC-PVP were efficient for seed coating. However, with increasing time (90days after seed coating), the viability of the strains decreased to 5.90 log CFU g-1 on the seed surface. Coated seeds showed resistance against disease and reduced disease incidence to even 0.00% compared to non-coated seeds under pot experiment. This study provides insights into the potential of seed coating to suppress fungal pathogens.

Drought is one of the major environmental stressors that significantly limits the germination, seedling growth, and productivity of cotton, particularly in arid and semi-arid regions. Recently, nanopriming has emerged as a novel and environmentally friendly seed treatment strategy to enhance plant resilience to drought stress. This study investigated the efficacy of nanopriming with Fe3O4 and ZnO nanoparticles (NPs) incorporated into cellulose microfibres (CMF) in improving drought tolerance of cotton (Gossypium hirsutum L.) under osmotic stress induced by polyethylene glycol (PEG). CMF extracted from cotton stalk residues was used as a sustainable carrier for Fe3O4 and ZnO NPs, yielding Fe/CMF and Zn/CMF composites. Cotton seeds were primed with Fe/CMF (50 and 100 ppm), Zn/CMF (150 and 200 ppm), and a non-primed control, and exposed to three water regimes (0%, 10%, and 15% PEG) to simulate increasing drought severity during germination and early seedling establishment. A comprehensive set of germination, growth, physiological, and biochemical traits was evaluated, including germination percentage, mean germination time, seedling vigour index, root and shoot growth, relative water content (RWC), proline accumulation, oxidative stress markers (malondialdehyde and hydrogen peroxide), and antioxidant enzyme activities (SOD, POD, and CAT). Nanopriming, particularly with 50 ppm Fe/CMF and 200 ppm Zn/CMF, significantly enhanced germination performance, seedling vigour, and biomass accumulation under moderate and severe PEG-induced drought stress. Moreover, nanoprimed seedlings maintained higher RWC, exhibited strongly upregulated antioxidant enzyme activities, accumulated greater proline levels, and showed marked reductions in lipid peroxidation and hydrogen peroxide content, indicating improved osmotic adjustment, redox homeostasis, and membrane stability under drought conditions. These results demonstrate that CMF-mediated delivery of Fe3O4 and ZnO NPs effectively mitigates drought-induced physiological and oxidative constraints during early cotton establishment, highlighting Fe/CMF and Zn/CMF nanopriming as a promising, sustainable strategy for enhancing early-stage drought tolerance in cotton-based production systems.

Low Bt toxin concentration in seeds results in low insecticidal efficacy in transgenic Bt cotton. In order to improve the insecticidal efficacy of seeds, two treatments with different amino acid combinations (5 amino acids comprising aspartic acid, glutamic acid, proline, methionine, and arginine; and 21 amino acids) were applied to two Bt cotton cultivars at peak boll stages in 2021 and 2022. The results showed that the amino acid treatments enhanced the seeds’ Bt toxin concentration by 13.5–34.2% compared with the untreated control in a two-year study. However, the difference for the Bt toxin was not significant between the two amino acid treatments. In the seeds, Bt toxin levels correlated positively with amino acid and soluble protein contents, as well as Glutamic-Pyruvic Transaminase (GPT) and Glutamate Oxaloacetate Transaminase (GOT) activities. Conversely, negative correlations were observed between the Bt toxin and the activities of protease and peptidase. Compared with the control, hazard boll rates were also reduced following application of the two amino acid combinations, while no difference was observed between the two amino acid treatments. Because the two treatments performed similarly, these results suggest that applying a simpler combination of five amino acids is an effective and efficient strategy for enhancing the insecticidal efficacy of cotton seeds.

Cotton is a vital cash crop in India that is predominantly cultivated in rainfed regions and is increasingly affected by climate change, which leads to poor seed germination and seedling establishment. To address these challenges and support sustainable agriculture, the current investigation aimed to develop and optimize bioencapsulated cotton seeds using plant growth-promoting bacteria (PGPB) to enhance seed germination, bacterial viability, seed shelf life, and crop yield. The bioencapsulation process was optimized using Response Surface Methodology via inverse gelation with sodium alginate, polyvinyl alcohol, starch, and glycerol. A Box-Behnken design was employed to evaluate germination percentage, seedling growth, bacterial survival, and seed vigour index. The optimal composition for bioencapsulation was 1.5% sodium alginate, 7.5% PVA, 5% starch, and 10% glycerol. Bioencapsulated seeds maintained > 80% seed germination and ~ 2.9 log₁₀ CFU/seed bacterial viability after 12months. Field studies conducted at two different locations in Punjab, India, showed that seeds bioencapsulated with B. thaonhiensis PAU_43RN and P. brassicacearum PAU_34RN improved seed cotton yield by 10.6% and 12.3%, and lint yield by 21.9% and 23.2%, respectively. This approach offers a promising solution for sustainable agriculture, reducing dependence on chemical fertilizers and enhancing resilience to climatic stresses.

The applicant Corteva Agriscience International Sàrl submitted a request to the competent national authority in Sweden to evaluate the confirmatory data that were identified for isoxaben in the framework of the maximum residue level (MRL) review under Article 12 of Regulation (EC) No 396/2005 as not available. To address the data gaps, a new analytical method for enforcement and its independent laboratory validation (ILV), plus additional studies on the magnitude of residues were submitted by the applicant. The outstanding data gaps were sufficiently addressed. The new information provided confirmed a no-residue situation in cotton seed and herbal infusions from flowers. The current MRL of 0.01 mg/kg for cotton seeds can therefore be confirmed. Since a more sensitive analytical enforcement method is now available, a lowering of the existing MRLs (0.05 mg/kg) for herbal infusions (flowers and roots) and hops to an limit of quantification (LOQ) of 0.01 mg/kg can be considered. The updated exposure calculations did not indicate a risk to consumers health.

Rapid, non-destructive and accurate prediction of cotton seed moisture content is important for assessing seed vigour and improving storage capacity. In this study, a prediction approach for cotton seed moisture content was developed based on machine learning (ML) and hyperspectral imaging. Using the cultivar Jinken 1161 as the experimental material, spectral data in the range of 935-1720nm were acquired. Outliers were removed using the Isolation Forest algorithm, and the samples were divided into calibration and prediction sets using the spectral-physicochemical value coordinate algorithm. The raw spectra were pre-processed using four methods, including Savitzky-Golay (SG) smoothing and standard normal variate transformation, before constructing traditional ML models [partial least square regression and multiple linear regression (MLR)] and deep learning (DL) models [convolutional neural network and long short-term memory network]. To reduce data redundancy and improve computational efficiency, feature wavelengths related to moisture content were selected using the successive projection algorithm and the least absolute shrinkage and selection operator (LASSO). Comparative analysis of different algorithmic combinations identified the optimal model, which was subsequently applied to hyperspectral images for pixel-wise prediction. This application enabled visualisation of the spatial distribution of moisture within individual cotton seeds. The results showed that, given the current sample size, ML models outperformed DL models. The SG-LASSO-MLR model achieved the best performance, with a prediction correlation coefficient (R2 p), root mean square error of prediction and residual predictive deviation of 0.9557, 0.908 and 4.77, respectively. These findings provide a feasible and effective technical solution for rapid and non-destructive detection of cotton seed moisture content. These outcomes offer valuable insights for seed quality evaluation and intelligent crop monitoring.

Chitosan nanoparticles induced germination and auxin-related gene regulations in cotton (Gossypium hirsutum L.) using random forest-based multi-output regression analysis

Cotton ( Cossypium ) fibres, which grow naturally in bolls around the seeds of cotton plants, have been used for centuries to produce fabric. The presumed natural function of cotton is that these lightweight and fluffy fibres may support wind dispersal of the seeds inside. However, cotton fibres are also frequently found in bird nests and we therefore argue that cotton fibres have also, or instead, evolved to encourage ‘caliochory': seed dispersal by animals that carry seeds to their nesting locations. To test this idea, we studied cotton collecting behaviour and use in nest building in two mousebird species (family Coliidae) in Eswatini. We revealed that cotton seeds are still present in large numbers inside the fibres that mousebirds used in their nests, and when the nests degrade after the breeding season, the seeds end up on the ground. By colour‐marking individual cotton seeds on their original vines, we found that seeds within a nest could originate from multiple parent plants located considerable distances from the nest (up to 1020 m). Moreover, video recordings revealed cases where mousebirds purposely collected cotton from other nests, potentially spreading the seeds even further away from the parent plant. Together, these findings show that cotton, by functioning as nest material, facilitates seed dispersal to favourable environments over barriers and ranges that might exceed distances possible by wind. We argue that the next step in the study of ‘caliochory' is to assess the ultimate benefits that both animal and plants obtain from seed dispersal through nest material, in order to determine whether cotton fibers have evolved specifically to facilitate seed dispersal.

Currently the cost and availability of protein-source feed for fattening of sheep is challenging. This study investigated different ratios of cotton seed cake with dried Sesbania sesban leaves on the feed intake, digestibility and growth performance of Semien sheep. The experiment was conducted for 90 days with 7 days of digestibility trials. A total of 20 intact male Semien lambs with an initial body weight of 15.16±0.96 were used. A randomized complete block design was used with five treatments: T1=100% cotton seed cake (CSC), T2=75% CSC+25% SSL, T3=50% CSC+50% SSL, T4=25% CSC+75% SSL and T5=100% SSL were offered 300g/day as feed basis, and the natural pasture hay and water were given ad libitum. Data on feed intake, body weight change and digestibility were subjected to analysis of variance (ANOVA) using statistical analysis system (SAS) 20.0. The lowest (p<0.01) hay dry matter (DM) intake was recorded for T1. The total DM intake was highly significant (p<0.01) among treatment groups in the order of T4>T3=T5>T2>T1. The apparent digestibility of DM, organic matter (OM) and crude protein (CP) was highly significant (p<0.001) among treatment groups. There were high records (p≤0.001) of body weight change at T3 and low in T1. The partial budget analysis showed that the net return of T3, T2, T1, T4 and T5 was 1024, 990.37, 903 and 790.12 ETB/treatment, respectively. The findings showed that from biological and economic responses, sheep fed on 50% CSC+50% SSL could be recommended.

Cotton seeds oils have their potential biological activity and specific physicochemical properties, due to which they are playing vital role in human nutritional diet for health benefits. The aim of this study was to compare secondary metabolites, physicochemical properties and fatty acids composition of seeds oil extracts of Gossypium hirsutum L. and Gossypium barbadense L. Cotton seeds used in this study were obtained from the Agricultural Research and Technology Corporation (ARTC), Wad Medani, Sudan. Seed oils methanolic extracts of Gossypium hirsutum and Gossypium barbadense were screened for their accumulated secondary compounds. Results indicated that G. hirsutum seed oil has more alkaloid than G. barbadense, whereas no variation in the amount of saponins, tannins, phenolic compounds, flavonoids and steroids &amp; terpenoids in two mentioned species. Physicochemical properties analyses were carried out using standard analytical methods. Results of physical properties showed that the specific gravity and refractive index@25C0 were 6.73 and 1.470 for G. hirsutum compared to 6.87 and 1.475 for G. barbadense respectively. Density (g/c3) of seed oils in order were 0.935 and 0.939 whereas pH was 6.76 and 5.76. The chemical properties of oils of two studied species recorded acid values (mg KOH/ g), % of free fatty acids, peroxide values (meq/Kg fat), saponification value (mg KOH/g) and iodine value (I/g oil) were in order 7.1and 20.3; 3.3and 10.3; 0.00 and 0.00; 188.0 and 195.0; 113.6 and 110.4 for G. hirsutum and G. barbadense respectively. Fatty acids composition of seed oils of two species were revealed using the gas chromatography-mass spectrometry (GC-MS) technique. The dominant unsaturated fatty acids of G. hirsutum seed oils were oleic acid 13.11% and linoleic acid 44.25%, while the saturated fatty acids were palmitic acid 29.16 %, methyl stearate acid (stearic acid) 5.06%, methyl tetradecanoate (methyl myristate) 1.48% and arachidic acid 0.67%. Comparatively, the unsaturated fatty acids of G. barbadense were oleic acid 22.63 % and linoleic acid 9.65 %, whereas the saturated fatty acids were palmitic acid 40.12%, methyl stearate (stearic acid) 7.18%, methyl tetradecanote (methyl myristate) 1,71% and arachidic acid 1.51%. In conclusion, the study revealed that oils extracted from G. hirsutum and G. barbadense, grown in Sudan contain the main saturated and unsaturated fatty acids. Physicochemical properties showed standard values in most parameters especially for G. hirsutum. Secondary metabolites obtained indicated the presence of biological activities in the oils of these species.

Seed aging is a complex biological process, the deterioration of oil crop seeds, in particular, has caused yield decline and economic loss to a large extent. However, research on the aging mechanism of oilseeds, such as cotton seeds, is still unclear. In this study, the physiological and biochemical changes in artificially aged cotton seeds were examined to further reveal the mechanism of seed aging and deterioration. Cotton seeds of “Xinluzao 74” were treated by artificial aging treatment method with high temperature (45°C) and high relative humidity (100%) for 1, 2, 3, 4, or 5 days, respectively, and untreated cotton seeds were used as control (CK). Our results showed that the germination rate, seed embryo viability, dehydrogenase activity, the activities of dehydrogenase antioxidant enzymes (SOD and POD), seed respiration rate, ATP content, ATP synthase activity and ATP synthase subunit mRNA integrity all showed a significant downward trend with the aging treatment time (P < 0.05), whereas the ROS generation (H2O2 content and ‧O2- production rate), relative conductivity, MDA content of seeds increased significantly, and the ultrastructure of cell membrane, mitochondria and chromatin of seed embryo was seriously damaged. Correlation analysis showed that there was a strong negative correlation between germination rate, SOD and POD activities, and respiration rate with H2O2 content (P < 0.05). This study reveals that excessive ROS, particularly H₂O₂, causes oxidative damage to ATP synthase subunit mRNAs, leading to impaired mitochondrial respiration and reduced seed vigor. These findings provide new molecular evidence linking oxidative RNA damage with seed aging, which could inform seed quality evaluation and storage strategies.

The stability of seed quality traits is a key factor for developing reliable cotton cultivars with consistent performance across diverse environments. This study evaluated six seed traits (1000-seed weight, crude protein, oil, ash, crude fiber, and moisture) across five commercial upland cotton cultivars grown in four distinct environments in Greece. Field trials were conducted in ten independent commercial fields per cultivar within each environment, with four samples collected from each field. Seed quality traits were analyzed using recommended official methods, and the Stability Index (SI) was calculated for each trait to quantify consistency across environments and cultivars. Traits such as 1000-seed weight, oil, and crude protein exhibited more consistent expression across environments and cultivars, whereas crude fiber and moisture showed substantial variation in stability across environments and cultivar × environment combinations, reflecting strong genotype × environment effects. Evaluation of relationships between traits revealed significant positive associations between 1000-seed weight and oil, and between crude protein and ash, as well as negative associations between 1000-seed weight and moisture and between crude protein and oil, indicating potential trade-offs among seed traits. Principal component analysis further highlighted trait associations and variability patterns across genotypes and environments. Overall, the combined use of Stability Index assessments and multivariate analyses provides a robust descriptive framework for evaluating cultivar performance and supporting cultivar selection under diverse agro-ecological conditions.

Objective: The objective of this study is to investigate the phytotoxic effects of a textile effluent using cotton species, with the aim of enabling non-potable reuse in the irrigation of this crop. Theoretical Framework: The textile sector generates waste, organic pollutants, and toxic substances, which, when released into water bodies, cause environmental imbalance. Phytotoxicity assesses the toxic effects of substances on plants, identifying the potentiation or inhibition of seed germination. Method: Acute phytotoxicity was conducted in the laboratory, with incubation for 120 hours, using Petri dishes and filter paper. Chronic bioassays were performed in an agricultural greenhouse for 21 days, at concentrations of 10 and 100 mg.L-1. The data were subjected to ANOVA, Dunnett and Tukey tests, and Germination Index. Results and Discussion: The results obtained revealed that in the acute bioassay there was an enhancement of cotton seed germination, with only some moderately phytotoxic indices obtained for the species *G. herbaceum*. In the chronic phytotoxicity test, no significant chronic effects on the plant growth and biomass endpoints were observed. Research Implications: It is suggested that future research should extend the plant life cycle in the chronic analysis and analyze the physiological aspects of cotton, as well as the soil. Originality/Value: This study contributes to the literature by reinforcing the potential of cotton seeds as resistant organisms in ecotoxicology and demonstrating that textile effluents may have the potential to stimulate plant development.

Abstract A novel copper(II) coordination complex, bis(2-hydroxybenzoate) bis(pyridine-3-carboxamide) copper(II) [Cu(HB) 2 (P3CA) 2 ], was synthesized and structurally characterized using single-crystal X-ray diffraction, spectroscopic techniques (FT-IR, UV–Vis), SEM–EDS, and XRF analyses. The complex exhibits a distorted octahedral geometry with N 2 O 4 coordination involving nitrogen from pyridine-3-carboxamide and oxygen from salicylate ligands. Hirshfeld surface analysis confirmed that strong hydrogen bonding and π–π stacking interactions stabilize the supramolecular structure. Computational studies using DFT revealed significant electron delocalization and donor-acceptor behaviour, suggesting high electronic mobility. Molar conductivity data highlighted efficient ionic dissociation in solution. The biological relevance was explored through phytotesting on cotton ( Gossypium hirsutum ) seeds, revealing that low concentrations of the complex significantly improved germination and seedling vigour. A concentration of 0.04 % showed 100 % germination and enhanced seedling biomass. These findings demonstrate the multifunctional nature of the complex, combining structural stability, electronic activity, and plant growth-promoting effects. The synthesized [Cu(HB) 2 (P3CA) 2 ] complex demonstrated significantly enhanced antimicrobial activity, 15–25 % against Bacillus subtilis and Pseudomonas aeruginosa , and 55 % against Candida albicans compared to salicylic acid, as evaluated by the agar well diffusion method.

Abstract Background The inhibition of cottonseed germination by soil salinization is a key obstacle affecting cotton production in Xinjiang. The most economical and effective measures to alleviate soil salinization damage are to explore the mechanism of cotton salt tolerance and to cultivate and popularize salt-tolerant varieties. Four cotton varieties were used in this study: the highly saline-alkali-resistant varieties Xinluzhong 82 and Xinluzhong 68 and the saline-alkali-sensitive varieties Xinluzhong 42 and Xinshi H12. These varieties were exposed to three different mixed saline-alkali concentrations: 0 mmol·L −1 (control, CK), 219 mmol·L −1 , and 365 mmol·L −1 . Results The germination characteristics, root traits, and changes in the storage material contents (crude fat, total protein, and total sugar) during cotton seed germination were analyzed. The results revealed that with increasing mixed saline-alkali concentrations, the relative water absorption, water absorption rate, germination rate, root length, diameter, area, and volume, and total sugar content of cotton decreased by 22.7%–90.1% on average, whereas the crude fat and protein concentrations increased by 7.6% on average. Under saline-alkali stress, cotton seeds undergo metabolic reallocation characterized by a decrease in carbohydrate content and the accumulation of lipids/proteins, potentially mitigating energy deficits and structural damage. Conclusions Under saline-alkali stress, highly resistant cotton varieties presented increased growth and adaptation during early root emergence. This was achieved by increasing water uptake and adjusting the root diameter. Under 365 mmol·L −1 saline-alkali stress, damages in these resistant varieties were correlated with their crude fat, protein, and storage substance contents. Changes in stored nutrients likely support cotton growth under stress. These findings reveal the physiological mechanisms underlying saline-alkali resistance in cotton and provide a scientific basis for the breeding of tolerant varieties. This study identified the key factors affecting cottonseed germination and offered a new direction for breeding programs.

Seed aging is a complex biological process, the deterioration of oil crop seeds, in particular, has caused yield decline and economic loss to a large extent. However, research on the aging mechanism of oilseeds, such as cotton seeds, is still unclear. In this study, the physiological and biochemical changes in artificially aged cotton seeds were examined to further reveal the mechanism of seed aging and deterioration. Cotton seeds of "Xinluzao 74" were treated by artificial aging treatment method with high temperature (45°C) and high relative humidity (100%) for 1, 2, 3, 4, or 5 days, respectively, and untreated cotton seeds were used as control (CK). Our results showed that the germination rate, seed embryo viability, dehydrogenase activity, the activities of dehydrogenase antioxidant enzymes (SOD and POD), seed respiration rate, ATP content, ATP synthase activity and ATP synthase subunit mRNA integrity all showed a significant downward trend with the aging treatment time (P < 0.05), whereas the ROS generation (H2O2 content and ‧O2- production rate), relative conductivity, MDA content of seeds increased significantly, and the ultrastructure of cell membrane, mitochondria and chromatin of seed embryo was seriously damaged. Correlation analysis showed that there was a strong negative correlation between germination rate, SOD and POD activities, and respiration rate with H2O2 content (P < 0.05). This study reveals that excessive ROS, particularly H₂O₂, causes oxidative damage to ATP synthase subunit mRNAs, leading to impaired mitochondrial respiration and reduced seed vigor. These findings provide new molecular evidence linking oxidative RNA damage with seed aging, which could inform seed quality evaluation and storage strategies.

Background: The use of plant-based feeds is increasing in the aquaculture industry. However, these diets have negative effects on fish, which must be addressed. Organic acids (OA) showed benefits in plant-based feed in aquaculture. Objectives: This project aimed to investigate the effect of adding cottonseed (CS) meal and a mixture of OA to the diet on blood immunity indices, histopathology, and gut microbial population in rainbow trout. Methods: To this end, 6 treatments consisting of 6 dietary formulations with 3 replications were designed in a 3×2 factorial arrangement. In this design, two levels of CS meal (0% and 15%) and three levels of the organic acid mixture (a mixture of lactic acid, citric acid, and potassium sorbate in equal proportions) at 0%, 0.5%, and 1% were added to the diet. A total of 270 rainbow trout with an Mean±SD weight of 0.14±0.35 g were stored in 18 aquariums containing 40 L of water, with a density of 15 fish per aquarium. The diets mentioned above were provided to the fish for 8 weeks at a daily rate of 3% to 4% of their biomass. After 8 weeks of rearing, blood and liver samples were collected from all treatments. Results: CS meal and OA had no significant effect on white blood cells, total immunoglobulin levels, or intestinal histology. CS meal did not significantly affect lysozyme, complement, total protein, albumin, or plasma globulin. However, the OA led to a significant increase in these parameters. The addition of CS meal increased lactic acid bacteria (LAB) and Vibrio sp. in the fish’s gut but did not affect the total number of gut bacteria. OA resulted in a decrease in total bacteria and Vibrio abundance while increasing LAB abundance in the gut. Conclusion: In conclusion, adding 15% CS meal to the diet of rainbow trout does not impact blood immune responses or intestinal tissue structure. However, it increased both beneficial (lactic acid bacteria) and harmful (Vibrio sp.) gut populations. On the other hand, adding OA effectively addresses this issue by increasing LAB populations and decreasing Vibrio sp. population. Furthermore, regardless of the addition of CS meal to the diet, incorporating OA improves innate immune parameters in the blood.

Contribution of oilseed waste on the development of the edible insect species Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae): From trash to cash.

IntroductionRapid and uniformgermination of cotton seeds is a prerequisite for achieving high and stable yields. Elucidating the molecular mechanisms underlying seed germination will help establish a genetic foundation for improving seed vigor, which is a key agronomic trait.MethodsIn this study, seeds from 259 Gossypium hirsutum germplasm accessions grown in seven environments over two years were evaluated for eight seed-vigor-related traits, including germination potential, germination rate, germination index, and vigor index. These eight traits exhibited abundant phenotypic variation within the population and were strongly correlated with each other. Correlation analysis, principal component analysis, and random forest importance evaluation identified the germination index, germination potential, and vigor index as the three traits most significantly and closely associated with seed vigor. Therefore, these three traits were used for genome-wide association analysis to identify important loci and genes controlling seed germination vigor in cotton.Results and discussionUsing a mixed linear model (MLM), six stable SNPs associated with the germination index (GI), three with germination potential (GP), and six with vigor index (VI) were identified, yielding a total of 15 stably associated SNPs across the three traits in multiple environments. Among them, locus A09:47545163 was repeatedly detected for both the germination index and potential. Using a 500-kb interval as the reference distance for candidate region delineation, 166 candidate genes harboring nonsynonymous mutations were identified. Further integrating GO/KEGG enrichment analysis, genotype effects at significant loci, and gene functional annotation, we prioritized 10 candidate genes that are potentially involved in the formation and regulation of seed vigor in cotton. Meanwhile, public TM-1 transcriptomic expression data provided exploratory supporting evidence for the functional inference of some candidate genes. These findings provide new evidence for elucidating the genetic basis of seed vigor in upland cotton and offer valuable gene resources for the development of high-vigor germplasm and related molecular markers.