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Macroalgae plays a significant role in the formulation of innovative and environmentally sustainable approaches to address food challenges. Specifically, green macroalgae serve as dietary supplements aimed at improving the health, growth, and feeding efficiency of various species of marine and freshwater fishes, as well as mollusks. The effects of Chaetomorpha linum extract (CLE) on growth performance, physiological responses, and disease resistance are studied in Bellamya bengalensis against Aeromonas hydrophila. In this experiment, adult B. bengalensis (4412 ± 165.25 mg) were randomly divided into 15 rectangular glass aquariums (35 snail/aquaria; 45 L capacity) and their basal diet was supplemented with different levels of CLE, including 0 (CLE0), 1 (CLE1), 2 (CLE2), 3 (CLE3), and 4 (CLE4) g/kg for 60 days. The growth performance in the CLE3 dietary group was significantly higher that of the CLE0 group, exhibiting both linear and quadratic trends in relation to dietary CLE levels (p < 0.05). The activities of pepsin, amylase, and lipase were found to be highest in CLE3 and lowest in CLE0. Both linear and quadratic responses to dietary CLE levels in digestive enzymes were observed (p < 0.05). The activities of superoxide dismutase and catalase in the hepatopancreas were found to be elevated in snails due to the synergistic effect of the supplemented CLE diet. Among different levels of diet given, CLE2-supplemented snails showed an increase in their enzyme activity (p < 0.05). Interestingly, all the CLE-treated snails expressed elevated levels of mucus lysozyme and mucus protein when compared to control (p < 0.05). Additionally, hepatopancreatic acid phosphatase and alkaline phosphatase activity were elevated in snails consuming CLE3 (p < 0.05). The transcription levels of immune-related genes, including mucin-5ac and cytochrome, were significantly elevated in snails that were fed a diet supplemented with 2–4 g of CLE/kg. Furthermore, the transcription level of the acid phosphatase-like 7 protein gene also increased in snails receiving CLE-supplemented diets. After a 14-day period of infection, snails that consumed a diet supplemented with 3–4 g/kg of CLE exhibited a notable increase in survival rates against virulent A. hydrophila. Based on the above findings, it is suggested that a diet supplemented with 3 g/kg of CLE may enhance growth, antioxidant and immune defense, and disease resistance in the freshwater snail B. bengalensis. Full article

Background/Objectives: Specific pharmacological osteoporosis therapy (SPOT) is regarded as a key intervention to reduce fracture risk and improve musculoskeletal function. Real-life data, particularly regarding functional muscular outcomes and pain trajectories, remain limited. This study aimed to longitudinally analyze bone mineral density, laboratory parameters, handgrip strength, functional performance, and pain symptoms under guideline-based SPOT. Methods: In this monocentric prospective real-life observational study, 178 patients (80.9% women; median age 82 years) with confirmed osteoporosis were followed for a median of four years. All patients received guideline-recommended antiresorptive or osteoanabolic therapy. Analyses included T-scores, 25(OH)D, calcium, handgrip strength, Chair Rise Test (CRT), tandem stance (TS), pain parameters, alkaline phosphatase (AP), HbA1c, fractures, comorbidities, and body mass index (BMI). Time-dependent changes were evaluated using linear mixed-effects models. Results: Bone mineral density improved highly significantly (ΔT-score ≈ +0.45 SD; p < 0.001), with no differences between therapy groups (antiresorptive vs. osteoanabolic) or BMI categories. Serum 25(OH)D levels increased markedly (Δ ≈ +20 nmol/L; p < 0.001), while calcium levels showed a small but highly significant decrease (Δ ≈ −0.047 mmol/L; p < 0.001), particularly under antiresorptive treatment. Dominant (Δ ≈ −1.95 kg; p < 0.001) and non-dominant handgrip strength (Δ ≈ −0.83 kg; p = 0.046) decreased significantly. In contrast, functional performance improved significantly: CRT time decreased by ~1 s (p = 0.004), and TS time increased by ~1 s (p = 0.007). Back pain decreased highly significantly (Δ ≈ −1.5 NRS; p < 0.001), while pain-free walking time (Δ ≈ +38 min; p = 0.031) and pain-free standing time (Δ ≈ +31 min; p = 0.038) both increased significantly. AP levels decreased significantly (p = 0.003), particularly among normal-weight patients. HbA1c changes were not significant. Overall, 73% of patients had at least one major osteoporotic fracture. Conclusions: In this real-life cohort, guideline-based specific pharmacological osteoporosis therapy was associated with significant improvements in bone mineral density, vitamin D status, functional performance, and pain-related outcomes. Despite a moderate decline in handgrip strength, balance- and mobility-related functional parameters improved, suggesting preserved or even enhanced functional capacity in daily life. These findings provide real-world evidence on the associations between SPOT, laboratory parameters, functional performance, and pain outcomes in a very elderly and multimorbid population. Full article

Background: Depression and Metabolic Dysfunction-Associated Steatotic Fatty Liver Disease (MASLD) are common chronic diseases, respectively. However, the causal and molecular links between them remain unclear. In order to explore whether depression contributes to an increased risk of MASLD and whether inflammation mediates this effect, we integrated multi-level evidence from the epidemiology of the National Health and Nutrition Examination Survey (NHANES), the genetics of GWAS, the transcriptomes of GEO, and single-cell RNA sequencing datasets. Methods: A multi-level integrative analysis strategy was used to validate this pathway. First, a cross-sectional epidemiological analysis based on NHANES data was used to reveal the association between depression and MASLD, and to explore the mediating role of inflammation and liver injury markers. Secondly, a two-sample Mendelian randomization analysis was used to infer the causal direction of depression and MASLD, and to verify the mediating effect of systemic inflammation and liver injury indicators at the genetic level. Then, the transcriptome co-expression network analysis and machine learning were used to screen the common hub genes connecting the two diseases. Finally, single-cell transcriptome data were used to characterize the dynamic expression of potential key genes during disease progression at cellular resolution. Results: Depression significantly increased the risk of MASLD, especially in women (OR = 1.39, 95%CI [1.17–1.65]). Parallel mediation analysis showed that high-sensitivity C-reactive protein (hs-CRP) (p < 0.001), γ-glutamyltransferase (GGT) (p < 0.001), and alkaline phosphatase (ALP) (p < 0.001) mediated this relationship. Mendelian randomization analysis confirmed the unidirectional causal effect of depression on MASLD, and there was no reverse association (β = 0.483, SE = 0.146, p = 0.001). Weighted gene co-expression network analysis and machine learning identified CD40LG as a potential molecular bridge between depression-associated immune modules and MASLD. In addition, single-cell data analysis revealed a stage-specific trend of CD40LG expression in CD4⁺ T cells during MASLD progression, while its receptor CD40 was also activated in B cells. In the female sample, CD40LG maintained an upward trend. However, the stability of this result is limited by the limited sample size. Conclusions: This study provides converging multi-omics evidence that depression plays a causal role in MASLD through inflammation-mediated immune signaling. The CD40LG-CD40 axis has emerged as an immune mechanism that transposes depression into the pathogenesis of MASLD, providing a potential target for the intervention of gender-specific metabolic liver disease. Full article

This study was focused on the assessment of fungal occurrence, mycotoxin dynamics, aflatoxin carry-over, and associated biochemical responses in dairy cattle. Moisture emerged as the dominant factor for fungal communities, promoting the co-proliferation of fungal genera adapted to high water activity conditions (a_w > 0.90) and antagonism against xerotolerant and xerophilic species. Aspergillus spp. dominated dry substrates (a_w < 0.75), Fusarium spp. showed strong positive associations with high-moisture matrices (a_w > 0.90), and Penicillium spp. exhibited intermediate, substrate-dependent behavior. Mycotoxin levels fluctuated non-linearly, independently of fungal counts: ochratoxin A (OTA) concentrations in corn silage increased from approximately 12 μg/kg at the onset of the ensiling period to >240 μg/kg at silo opening, indicating dynamic mycotoxin accumulation during storage, while zearalenone (ZEA) oscillated from 40 to 170 µg/kg. Despite the variation in total aflatoxins (AFLA-T) across feed matrices, aflatoxin M1 (AFM1) in milk remained low (0.0020–0.0093 μg/kg), confirming limited carry-over. Serum biochemical parameters—alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), alkaline phosphatase (ALP), total bilirubin (BIL-T), total protein (PROT-T)—remained within physiological limits, yet multivariate analyses revealed metabolic modulation linked to aflatoxin exposure. AFM1 explained >7% of the variance in serum biochemical profiles according to PERMANOVA (p = 0.002), showed significant MANOVA effect (Pillai = 0.198), and displayed a significant canonical association (p < 10⁻¹³). Linear discriminant analysis further separated Normal vs. Borderline hepatic profiles, indicating subclinical physiological adaptation to chronic low-dose exposure. Full article

Plastic pollution is acknowledged as a serious problem for ecosystems. Among these plastics, polystyrene nanoplastics (PS-NPs) are emerging environmental pollutants, and their biological effects on hepatotoxicity are the least explored. Therefore, the present work examined the effect of PS-NPs on the hepatic transcription of the antioxidant genes Hmox1 and Sod3 in mice (n = 6, treatment (PS-NPs) vs. vehicle group (Veh)), mediated by RORγ and epigenetic modifications. The results show that PS-NP mice had significantly reduced body weight; increased activity of adenosine triphosphate (ATP), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH), and Complexes I, III, and V in the liver; and increased Alanine Transaminase (ALT), Aspartate Transaminase (ASP), Alkaline Phosphatase (ALP), malondialdehyde (MDA) and reactive oxygen species (ROS) compared to the Veh group. Furthermore, PS-NPs resulted in considerably lower relative mRNA expression of Hmox1, Sod3, and RORγ in the liver than the Veh group. Likewise, when compared to Veh, PS-NPs significantly reduced the enrichment of RORγ, as well as the occupancies of the key components of the transcriptional activation pathway (P300, SRC1, Pol II, Ser5-Pol II, and Ser2-Pol II) at the loci of Hmox1 and Sod3. In comparison to Veh, PS-NPs showed downregulated occupancies of the histone active marks H3K9ac and H3K18ac, while H3K4me3 and H3K27me3 were higher at the target loci of Hmox1 and Sod3. In conclusion, the present study highlights that PS-NPs induce oxidative stress by modifying Hmox1 and Sod3 in mice’s livers through histone changes and nuclear receptor RORγ modulation. Full article

A global interest in the cultivation of Kalibaus (Labeo calbasu) has emerged due to decreasing natural stocks and a consistent rise in market value and demand. Given these concerns, understanding the species’ physiological responses to environmental changes is crucial. The present research aimed to assess the effect of varying environmental temperatures on metabolism, haemato-biochemical indices, hormonal concentrations and immune responses in L. calbasu. This study was conducted in triplicate using 100 L glass aquariums at four different temperatures: 22, 26, 30, and 34 °C. The highest weight and length gain were observed at 30 °C, while the lowest occurred at 22 °C. Notably, the best feed conversion ratio (FCR) of 1.51 ± 0.03 was also recorded at 30 °C. Although haematological and biochemical parameters remained within normal ranges, they varied with temperature changes. Indicators of cold and heat stress were evident through lower hematocrit levels and higher white blood cell (WBC) counts. Biochemical indicators such as serum albumin (1.84 ± 0.05 g dL⁻¹), serum globulin (1.64 ± 0.06 gdL⁻¹), HCO₃ (30.93 ± 0.62), Na⁺ (115.60 ± 3.72 mmolL⁻¹), alkaline phosphatase (93.33 ± 9.39 AP, IUL⁻¹), and AST/SGOT (21.00 ± 4.55 UL⁻¹) were significantly higher at 30 °C. Regarding hormonal responses, peak levels of growth hormone (GH), triiodothyronine (T3) (1.44 ± 0.07 ngmL⁻¹), and thyroxine (T4) were recorded at 30 °C. Meanwhile, serum cortisol (1.62 ± 0.06 µgdL⁻¹) and adrenocorticotropic hormone (ACTH) (18.01 ± 3.26 pgmL⁻¹) were highest at 34 °C. Immune responses were strongest between 26 and 30 °C. In conclusion, the results suggest that L. calbasu should ideally be cultured between 26 and 30 °C for optimum growth and health, making it ideal for commercial farming. Full article

Black spot disease substantially impairs both the aesthetic quality and commercial viability of affected Pingguoli pears. Previous studies have shown that Alternaria alternata and A. tenuissima are the pathogens that cause black spot disease. Essential oils represent novel alternatives to synthetic fungicides to control these pathogens. This study extracted Artemisia sieversiana essential oil (AsEO) by hydro-distillation using a crystal tower pure dew essential oil machine. The chemical compositions of AsEO were analyzed via gas chromatography–mass spectrometry (GC–MS). A total of 42 compounds were detected. 1,8-cineole, trans-caryophyllene, (1R,4S)-1,7,7-trimethylbicyclo [2.2.1] heptan-2-yl acetate, (±)-camphor, and β-myrcene were identified as the five main constituents. Moreover, the antifungal activity of AsEO was assessed against black spot on Yanbian Pingguoli pear in China. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values were determined as 0.10% (v/v) and 0.12% (v/v), respectively. Scanning electron microscopy (SEM) analysis revealed that treatment with AsEO induced significant morphological aberrations in A. alternata and A. tenuissima mycelia, including surface roughening, hyphal collapse, and loss of structural integrity. Concurrently, a marked increase in alkaline phosphatase (AKP) enzyme activity and electrical conductivity was observed, a key indicator of cell wall and plasma membrane permeabilization and damage. When the concentration of AsEO was less than 120 µg/mL, there was no toxicity to keratinocytes (HaCaTs) and skin fibroblasts (NHSFs). In summary, this study provides a theoretical basis for the development of AsEO as a fungicide against black spot disease on Pingguoli pear in China. Full article

Ozone (O₃) is a reactive oxidant increasingly applied in biomedical settings, yet its dose-dependent effects on innate immune cells, particularly those from non-human species, remain insufficiently defined. Within a One Health framework, this study examined how two clinically relevant O₃ exposure regimens (30 µg/mL and 90 µg/mL) affect porcine neutrophils and monocytes isolated from peripheral blood. Cell viability, reactive oxygen and nitrogen species (RONS) production, and the activity of key enzymes (myeloperoxidase, elastase, alkaline phosphatase, arginase) were assessed at 1 h and 24 h post-exposure. The lower dose induced mild functional activation without compromising viability, whereas the higher dose triggered pronounced oxidative stress, enhanced degranulation, and reduced neutrophil viability by more than 60%. Neutrophils exhibited a stronger and more dynamic response than monocytes, which retained viability and differentiation capacity at 30 µg/mL but showed impaired function at 90 µg/mL. These findings highlight the dual nature of O₃, where controlled exposure may support immunomodulation, while excessive dosing disrupts cell function. Defining safe and effective therapeutic windows remains critical for future applications. Full article

Improving phosphorus (P) utilization in broilers is crucial for reducing feed costs and environmental pollution. Bone mineralization trait is strongly associated with P utilization in poultry and is thus often used as an alternative trait for evaluating P utilization. Dentin matrix protein 1 (DMP1), an essential matrix protein for bone mineralization and P deposition, has been shown to be actively involved in P utilization in broilers, but the underlying mechanisms remain unclear. The current study aimed to investigate the possible mechanisms whereby DMP1 regulates P utilization of poultry by using gene silencing and overexpression technologies, combined with an in vitro model of primary broiler osteoblasts. The results showed that DMP1 overexpression augmented the P utilization of broiler osteoblasts, characterized by significant increases (p < 0.001) in P utilization rate, mineralization formation, alkaline phosphatase activity, and bone gla protein content. Meanwhile, DMP1 overexpression effectively (p < 0.05) activated the focal adhesion kinase (FAK) signaling, along with obvious (p < 0.01) decreases in fibroblast growth factor 23 (FGF23) expression and production. In contrast, DMP1 silencing reversed (p < 0.05) the above effects. Consistently, FAK activation promoted (p < 0.05) P utilization accompanied by remarkable (p < 0.05) decreases in FGF23 expression and production. Furthermore, gain- and loss-of-function assays demonstrated that a high level of FGF23 contributed to impaired P utilization, while a low level was beneficial. Interestingly, blocking FAK signaling not only recovered (p < 0.05) the FGF23 expression and production in DMP1 overexpressed cells but also obviously (p < 0.05) weakened their P utilization. These findings indicate that DMP1 inhibits FGF23 expression by activating FAK, thereby contributing to P utilization in broiler osteoblasts. They reveal a novel DMP1-FAK-FGF23 regulatory axis in broiler osteoblasts and provide a potential target for improving P efficiency in poultry. Full article

Cardiovascular disease (CVD) remains the leading cause of mortality among hemodialysis (HD) patients, underscoring the need for reliable biomarkers for early diagnosis and management. Serum intercellular adhesion molecule-1 (ICAM-1) has been investigated for years as a potential CVD marker but has yet to establish clinical utility. In a cohort of 142 HD patients, we examined the potential of serum ICAM-1 as a CVD biomarker and evaluated whether confounding factors, including low-grade inflammation and chronic kidney disease–mineral bone disorder (CKD-MBD), limit its diagnostic value. In addition to serum ICAM-1, routine biochemical parameters, bone alkaline phosphatase (bALP), and nitric oxide (NO) were measured. Serum levels of ICAM-1, bALP, and NO did not differ between patients with and without CVD, defined by a positive history of coronary heart disease, stroke, or peripheral arterial disease. Serum ICAM-1 concentrations were higher in HD patients with inflammation, as indicated by C-reactive protein levels >1 mg/dL. ICAM-1 showed no correlation with NO, a marker of endothelial dysfunction, but was positively correlated with bALP, a marker of CKD-MBD. In conclusion, serum ICAM-1 is not a reliable biomarker of CVD in HD patients. Its diagnostic utility appears confounded by inflammation and disturbances in bone turnover. Full article

The design of biomaterial scaffolds for bone tissue engineering requires a balance between bioactivity, porosity, mechanical stability, and osteoinductivity. Kappa- (KC) and iota-carrageenan (IC) have been explored for scaffold fabrication due to their biocompatibility and structural similarity to glycosaminoglycans. However, there are limited reports on how their distinct sulfation degree affects the osteogenic differentiation of cells cultured on them. While laponite has been reported as an osteoinductive nanoclay, its combined effect with different carrageenan types and its concentration-dependent effect on scaffold functionality remain unexplored. Therefore, we developed composite scaffolds comprising poly(vinyl alcohol) (PVA) and gelatin (GEL), reinforced with kappa- or iota-carrageenan (KC, IC) and functionalized with two different concentrations of laponite (LAP), 0.5 and 1% w/v, to monitor composition-structure-function relationships. The scaffolds were fabricated via lyophilization and dual crosslinking, and characterized for their physicochemical, structural, mechanical, and biological properties. The incorporation of both carrageenans into scaffolds, maintained high swelling ratios of 600% after 24 h, and increased porosity without altering their apparent density (0.09–0.11 g/cm³), whereas LAP preserved interconnectivity, densified pore walls, raised their compressive modulus at >220 kPa, and improved stability (>60% mass retained after 40 days). In vitro validation using MC3T3-E1 pre-osteoblastic cells demonstrated robust cytocompatibility, with the LAP-containing scaffolds significantly promoting cell adhesion, proliferation, and osteogenic differentiation, evidenced by elevated alkaline phosphatase activity, calcium production and collagen secretion. Direct comparison between KC and IC scaffolds confirmed that differences in sulfate substitution modulated scaffold stiffness, swelling, and degradation, while variation in LAP concentration affected the biological response, with the 0.5 wt% concentration favoring early cell proliferation, whereas the 1 wt% significantly promoted the osteogenic differentiation. This compositional strategy demonstrates how tuning the interplay between carrageenan and laponite can balance scaffold hydration, mechanical and biological properties, thereby guiding the design of scaffolds for bone repair. Full article

Probiotics hold great potential in aquaculture, as they can effectively modulate gut microbiota and improve fish health, thereby enhancing farming efficiency. Translating this potential into practical application critically relies on screening high-efficacy probiotic strains. This study evaluated the growth-promoting and health-enhancing effects of probiotic candidates Lactobacillus rhamnosus GG (LGG), Lactobacillus plantarum FZU310 (LP-FZU310) and Bacillus subtilis FZU103 (BS-FZU103) in largemouth bass (Micropterus salmoides). After feeding different probiotics for 30 days, the growth, antioxidant, and intestinal enzyme indicators of M. salmoides were detected. BS-FZU103 demonstrated superior efficacy among the tested strains, showing significant differences in both specific growth rate (SGR) (p < 0.05) and condition factor (CF) (p < 0.05). It also markedly enhanced hepatic antioxidant status, elevating superoxide dismutase and glutathione peroxidase activities while reducing malondialdehyde levels by 80%. Improved liver integrity was indicated by significant decreases in serum alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase. Digestively, BS-FZU103 specifically increased intestinal amylase activity by 14.7%, without affecting protease or lipase, suggesting enhanced carbohydrate digestion. 16S rRNA sequencing revealed BS-FZU103 remodeled gut microbiota, increasing Proteobacteria abundance at the phylum level and enriching Bacillus while reducing Clostridium sensu stricto 1 at the genus level. Functional prediction based on PICRUSt2 indicated an enhanced metabolic potential of the gut microbiota, with inferred upregulation of pathways related to carbohydrate transport and metabolism (e.g., ABC transporters) and intestinal enzymatic activities. Collectively, BS-FZU103 is associated with metabolic modulation, promoting M. salmoides growth through gut microbiota remodeling, hepatic antioxidant fortification, and targeted augmentation of carbohydrate utilization efficiency. Full article

Cadmium (Cd) is a typical pollutant with strong toxicity even at low concentrations. In the marine environment, Cd is a problem of magnitude and ecological significance due to its high toxicity and accumulation in living organisms. The clam Meretrix meretrix is a useful bioindicator species for evaluating heavy-metal stress. This study investigated the extent of recovery from Cd²⁺-induced oxidative and immune impairments in M. meretrix gills achieved by short-term depuration. Clams were exposed to 3 mg/L Cd²⁺ for six days or three days followed by three days of depuration, and the Cd contents, morphological structure, osmoregulation, oxidative stress, and immune responses in the gills were evaluated. The results showed that gill Cd contents increased with exposure, reaching 9.857 ± 0.074 mg·kg⁻¹ on day 3 but decreased slightly to 8.294 ± 0.056 mg·kg⁻¹ after depuration, while reaching 18.665 ± 0.040 mg·kg⁻¹ on day 6 after continuous exposure. Histological lesions, including lamellar fusion, hemolymphatic sinus dilation, and ciliary degeneration, partially recovered after depuration. Reactive oxygen species (ROS) and malondialdehyde (MDA) levels decreased significantly, while DNA-protein crosslinking rate (DPC) and protein carbonyl (PCO) showed minor reductions. Total antioxidant capacity (T-AOC) and the activities of Ca²⁺/Mg²⁺-ATPase (CMA), cytochrome c oxidase (COX), succinate dehydrogenase (SDH), and lactate dehydrogenase (LDH) increased by over 10% during depuration, though these changes were not statistically significant. Lysozyme (LZM) activity and MT transcript levels increased progressively with Cd exposure, indicating their suitability as biomarkers of Cd stress. Acid and alkaline phosphatase (ACP, AKP) activities and Hsp70 and Nrf2 mRNA transcripts exhibited inverted U-shaped response consistent with hormetic response. ACP and AKP activity levels rose by more than 20% after depuration, suggesting partial restoration of immune capacity. Overall, Cd exposure induced oxidative damage, metabolic disruption, and immune suppression in M. meretrix gills, yet short-term depuration allowed partial recovery. These findings enhance understanding of Cd toxicity and reversibility in marine bivalves and reinforce the usage of biochemical and molecular markers for monitoring Cd contamination and assessing depuration efficiency in aquaculture environments. Full article

Feeding Chinese mitten crabs with fresh-frozen fish causes nutritional imbalance and increases disease risk. Compound feed offers better nutrient balance but still requires improvements in palatability and growth performance. This study evaluated the effects of replacing fresh-frozen fish with glutamine dipeptide-supplemented formulated diet on growth, hepatopancreas health, and edible quality, aiming to inform feed formulation strategies. A five-month feeding trial (June–October) was conducted with two treatments: the experimental group received only glutamine dipeptide compound feed, while the control group was fed a mix of fresh-frozen fish and compound feed. Crabs in the experimental group showed significantly higher body weight, length, and width. No significant differences were found in the hepatopancreatic index, gonadosomatic index, meat yield, or total edible yield. In October, the experimental group showed lower malondialdehyde (MDA) levels in the hepatopancreas and higher alkaline phosphatase (AKP) and acid phosphatase (ACP) activities in males. In females, hemolymph AKP and ACP were higher in the control, while glutamic pyruvic transaminase (GPT) was higher in the experimental group. Whether this is related to a potential risk of liver damage or a reaction at a special stage remains to be further verified. Digestive enzyme activities (protease, lipase, amylase) were generally higher in the experimental group, particularly in August (p < 0.05). In October, protease activity in females and lipase activity in males were significantly higher than in controls (p < 0.05). Nitrogen and phosphorus retention in muscle was also significantly higher, indicating better nutrient utilization (p < 0.05). Overall, these findings indicate that a glutamine dipeptide-supplemented diet provides a more effective and sustainable alternative to fresh-frozen fish over a five-month rearing period, improving digestive physiology, feed efficiency, growth performance, and edible quality and flavor. Full article

Despite decades of structural and kinetic characterization, the full spectral molecular vibrations that accompany the catalysis in alkaline phosphatase (ALP) have remained largely unexplored. In this study, we combine in situ real-time attenuated total reflection Fourier transform infrared (ATR-FTIR) measurements over a large energy range to track the hydrolysis of p-nitrophenyl phosphate (PNPP) and inorganic phosphate (Pi) over a large range of enzyme concentrations. From the static spectra of the pure components (ALP, PNPP, PNP, Pi), we identify their characteristic vibrational frequencies and use them as reference points for the time-resolved spectra. The reaction reveals a monotonic growth of the inorganic-phosphate band at 1077 cm⁻¹. At the highest alkaline phosphatase concentration, we resolve two blue shifts in the nitro/aromatic region (1510 → 1518 cm⁻¹; 1494 → 1499 cm⁻¹), two red shifts in the fingerprint region (1345 → 1340 cm⁻¹; 1294 → 1290 cm⁻¹), and a splitting of the ~1592 cm⁻¹ band into 1595 and 1583 cm⁻¹. In conclusion, by anchoring the time-resolved spectra to the static spectra of individual constituents, we were able to resolve the infrared readout of the enzymatic reaction, offering a generalizable approach for FTIR-based tracking of catalytic processes. Full article