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18 pages, 2128 KB  
Article
Effects of Dietary Squalene Supplementation on the Growth Performance and Disease Resistance of Largemouth Bass
by Shan Liu, Mengmeng Chen, Yan Meng, Mingyang Xue, Yong Zhou, Liping Zhang, Peng Chen, Yuding Fan, Yazhen Yang and Zhenyu Huang
Vet. Sci. 2026, 13(5), 448; https://doi.org/10.3390/vetsci13050448 - 1 May 2026
Abstract
Feed supplements play a crucial role in improving and maintaining fish health in modern aquaculture practices. Squalene is a functional lipid naturally present in fatty tissues, possessing numerous beneficial biological properties and wide applications in the food and pharmaceutical industries. In this study, [...] Read more.
Feed supplements play a crucial role in improving and maintaining fish health in modern aquaculture practices. Squalene is a functional lipid naturally present in fatty tissues, possessing numerous beneficial biological properties and wide applications in the food and pharmaceutical industries. In this study, the effects of 100 mg/kg (S1), 200 mg/kg (S2), 300 mg/kg (S3), and 400 mg/kg (S4) of dietary squalene supplementation over four weeks on growth performance, antioxidation, hepatoprotection, hypoxia tolerance, immune relative genes expression, and disease resistance of largemouth bass (Micropterus salmoides) were assessed. The results showed that squalene supplementation significantly increased the weight gain rate (WGR) and specific growth rate (SGR) of largemouth bass (p < 0.05). Serum glucose (GLU) levels were significantly decreased in all squalene-supplemented groups (p < 0.05). Squalene supplementation had minimal effect on serum triglyceride (TG) and total cholesterol in (TCHO) levels. A decrease in malondialdehyde (MDA) level, but accompanied by increases in superoxide dismutase (SOD) and hepatic catalase (CAT) activities, was observed in the S1 group supplemented with squalene. These suggest that squalene may mitigate free radical damage and promote health in largemouth bass. Dietary squalene supplementation enhanced intestinal enzyme activities (trypsin, lipase, and α-amylase) in largemouth bass without inducing any apparent hepatic or histopathological alterations. Squalene supplementation improved hypoxia tolerance and antiviral gene expression (mx, ifn-γ, and irf3) while suppressing the expression of inflammatory cytokine (il-1β, il-8, and tnf-α). The survival rate following LMBRaV infection was significantly higher in the S1 group (100 mg/kg group) compared to the control (p < 0.05). In conclusion, this study demonstrated that adding squalene into the diet of largemouth bass at an optimal level of 100 mg/kg effectively promotes growth performance, enhances digestive enzyme activity and hypoxia tolerance, and modulates lipid metabolism and immune gene expression, thereby contributing to improved resistance against LMBRaV. These findings confirm that squalene can serve as a beneficial functional feed additive in aquaculture. Full article
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14 pages, 3941 KB  
Article
In Vitro TLR4 Stimulating Bioactivities of Amylase/Trypsin-Inhibitors from Wheat (Triticum aestivum L.) Bred from 1891 to 2010
by Manjusha Neerukonda, Sabrina Geisslitz, Darina Pronin, Valentina Curella, Sibylle Neufang, Sandra Koch, Klajdi Begaj, Ernesto Bockamp, Heiko Weichert, Andreas Börner, Hans Weber, Katharina Anne Scherf and Detlef Schuppan
Foods 2026, 15(9), 1541; https://doi.org/10.3390/foods15091541 - 29 Apr 2026
Viewed by 122
Abstract
Wheat amylase trypsin inhibitors (ATIs) are prominent allergens in Baker’s asthma and contribute to innate inflammation in non-celiac wheat sensitivity (NCWS), linking them to metabolic and autoimmune diseases. Their tetra-, di-, and monomeric forms, stabilized by disulfide bonds, confer resistance to digestion, baking, [...] Read more.
Wheat amylase trypsin inhibitors (ATIs) are prominent allergens in Baker’s asthma and contribute to innate inflammation in non-celiac wheat sensitivity (NCWS), linking them to metabolic and autoimmune diseases. Their tetra-, di-, and monomeric forms, stabilized by disulfide bonds, confer resistance to digestion, baking, and heating. Although proteomic studies reveal minor variation in ATI subtypes among cultivars and major variation among species, the influence of environment and wheat genotype on ATI levels and TLR4-stimulating activity remains unclear. We assessed the effect of the environment on the in vitro inflammatory bioactivity of ATIs extracted from 60 German wheat genotypes focusing on breeding over time between 1891 and 2010, and cultivation across three climatically distinct years. We found considerable genotype-dependent variation in ATI bioactivity that did not correlate with ATI subtype abundance, and observed no consistent difference between old and modern cultivars. ATIs from samples grown in 2019, a warm and dry year, showed reduced TLR4 activity, highlighting the significant impact of environmental conditions on inflammatory ATI bioactivity. Full article
(This article belongs to the Section Grain)
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34 pages, 1608 KB  
Article
Complementary and Synergistic Effects of Dietary Bacillus subtilis natto and Lactobacillus plantarum on Growth, Immunity and Intestinal Microflora Restructuring in Juvenile Japanese Flounder (Paralichthys olivaceus)
by Yukun Zhang, Manabu Ishikawa, Shunsuke Koshio, Saichiro Yokoyama, Na Jiang, Weilong Wang, Jiayi Chen, Noman Waheed and Xiaoxiao Zhang
Fishes 2026, 11(5), 259; https://doi.org/10.3390/fishes11050259 - 24 Apr 2026
Viewed by 134
Abstract
Prophylactic antibiotic use in intensive aquaculture promotes antimicrobial resistance, necessitating the development of microbial-based interventions. This study evaluated the individual, complementary, and synergistic effects of Bacillus subtilis natto (BSN) and Lactobacillus plantarum (LP) on the physiological performance and intestinal microecology of juvenile Japanese [...] Read more.
Prophylactic antibiotic use in intensive aquaculture promotes antimicrobial resistance, necessitating the development of microbial-based interventions. This study evaluated the individual, complementary, and synergistic effects of Bacillus subtilis natto (BSN) and Lactobacillus plantarum (LP) on the physiological performance and intestinal microecology of juvenile Japanese flounder (Paralichthys olivaceus). Over a 60-day trial, juveniles (initial weight: 5.81 ± 0.03 g) received a basal diet (CON) or a diet supplemented with 107 CFU/g of BSN, LP, or both (BSN+LP). The BSN+LP consortium elicited complementary improvements in final body weight (21.39 ± 0.75 g vs. 18.66 ± 0.44 g in CON) and feed conversion efficiency (p < 0.05). Transcriptomic analysis revealed synergistic upregulation of digestive proteases (trypsin, chymotrypsin). Notably, an in pro-inflammatory markers (IL-1β, TNF-α) was counterbalanced by substantial upregulation of anti-inflammatory cytokines (IL-10, 5.65-fold; TGF-β1, 4.48-fold), suggesting the induction of mucosal tolerance rather than pathological enteritis. High-throughput 16S rRNA sequencing showed that the control cohort had a potential baseline microbial, characterized by a high relative abundance of Proteobacteria. BSN+LP administration significantly altered this microbial community into a fermentative eubiosis enriched in Firmicutes and Bacteroidota. Correlation network analyses confirmed negative interaction dynamics: increased abundance of Lactobacillus, Bacteroides, and Muribaculaceae was negatively correlated with baseline pathobiont abundance. These findings indicate that co-administration of BSN and LP hypothetically enhances metabolic energy harvest via short-chain fatty acid-producing taxa, strengthens the gut–immune axis, and competitively mitigates opportunistic pathogens. Full article
17 pages, 412 KB  
Article
Functional Evaluation of Pomegranate (Punica granatum) Juice Byproducts as Dietary Additives in Red Seabream (Pagrus major): Effects on Growth Performance, Antioxidant Response, Immunity, and Resistance to Edwardsiella tarda
by Ki-Tae Kim, Tae Hoon Lee, Hwa Yong Oh, Da Ye Kang, Do Hyun Kwon, Young Wook Kim, Bo Seong Gu, Dona Thilini Udarika Samaraweera and Hee Sung Kim
Antioxidants 2026, 15(4), 517; https://doi.org/10.3390/antiox15040517 - 21 Apr 2026
Viewed by 251
Abstract
This study evaluated the potential of pomegranate (Punica granatum) juice byproducts (PJB) as a functional dietary additive for juvenile red seabream (Pagrus major). Four experimental diets were formulated to contain various levels of PJB (0, 2.5, 5.0, and 10.0 [...] Read more.
This study evaluated the potential of pomegranate (Punica granatum) juice byproducts (PJB) as a functional dietary additive for juvenile red seabream (Pagrus major). Four experimental diets were formulated to contain various levels of PJB (0, 2.5, 5.0, and 10.0 g/kg) and fed to fish with an initial body weight of 7.0 ± 0.01 g for 8 weeks. Growth performance, feed utilization, digestive enzyme activity, whole-body composition, plasma biochemical parameters, antioxidant responses, immune parameters, and resistance to Edwardsiella tarda infection were evaluated. Fish fed the diet containing 2.5 g/kg PJB exhibited significantly higher final body weight, weight gain, and specific growth rate compared with the control group and those with higher PJB doses, whereas feed intake, feed efficiency, and protein efficiency ratio were not significantly affected by dietary treatment. Intestinal trypsin and lipase activities were significantly elevated in the PJB2.5 group, whereas amylase activity remained unchanged. Whole-body proximate composition and plasma biochemical parameters, including aspartate aminotransferase, alanine aminotransferase, total cholesterol, glucose, and total protein, were not significantly influenced by dietary PJB supplementation. Dietary inclusion of PJB at 2.5 g/kg also significantly enhanced plasma antioxidant enzyme activities, as evidenced by increased superoxide dismutase and glutathione levels, while catalase activity was elevated in fish fed the PJB2.5 and PJB5 diets. Innate immune responses were also stimulated, with significantly higher serum lysozyme activity and interleukin-1 levels observed in fish fed the PJB2.5 diet. Following experimental challenge with E. tarda, fish fed diets containing 2.5 and 5.0 g/kg PJB exhibited significantly higher cumulative survival than the control group. In conclusion, dietary supplementation with PJB at 2.5 g/kg improved growth performance, digestive capacity, antioxidant status, innate immune responses, and disease resistance in juvenile P. major without adverse physiological effects. Full article
(This article belongs to the Special Issue Natural Antioxidants and Aquatic Animal Health—3rd Edition)
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22 pages, 2527 KB  
Article
A Degradable Nanosystem Based on Small Gold Nanoparticles and Albumin for Amyloid Aggregation Inhibition
by Matías Levio, Francisco Rossel Carrera, Fredys Sánchez Hoyos, Maycol Huerta, Carlos Alamos, Rodrigo Vásquez-Contreras, Marcelo J. Kogan and Eyleen Araya Fuentes
Pharmaceutics 2026, 18(4), 504; https://doi.org/10.3390/pharmaceutics18040504 - 19 Apr 2026
Viewed by 448
Abstract
Background/Objectives: Beta amyloid (Aβ) aggregates play a central role in the pathophysiology of Alzheimer’s disease (AD), and their detection and modulation remain major challenges in developing effective therapeutic and diagnostic strategies. Previously, gold nanoparticles with plasmonic and optical properties in the near-infrared [...] Read more.
Background/Objectives: Beta amyloid (Aβ) aggregates play a central role in the pathophysiology of Alzheimer’s disease (AD), and their detection and modulation remain major challenges in developing effective therapeutic and diagnostic strategies. Previously, gold nanoparticles with plasmonic and optical properties in the near-infrared (NIR) region and photothermal capabilities have been designed for detecting and disaggregating Aβ aggregates. However, these systems often face limitations related to biodegradability, long-term accumulation, and safety. In this work, a degradable NIR-responsive nanosystem based on small gold nanoparticles (sAuNPs), potentially excretable due to their small size, encapsulated within bovine serum albumin (BSA) and functionalized with the all-D peptide D3, was developed to inhibit Aβ aggregation. Methods: sAuNPs (~5–6 nm), functionalized with HS-PEG-NH2, were encapsulated into BSA nanoparticles using a desolvation method and subsequently conjugated to D3, resulting in the nanosystem f-sAuNPs-BSANPs-D3. The nanosystem was characterized by UV–Vis–NIR spectroscopy, dynamic light scattering, zeta potential analysis, electron microscopy, and nanoparticle tracking analysis. The effects of the nanosystem on Aβ1–42 aggregation were evaluated using a thioflavin T assay and electron microscopy. Additionally, the effects of f-sAuNPs-BSANPs-D3 on cell viability and its stability against trypsin digestion were assessed. Results: The nanosystem exhibited a measurable photothermal response under NIR irradiation and significantly reduced fibril formation. It did not affect the viability of SH-SY5Y neuronal cells at the tested concentrations. Trypsin incubation experiments demonstrated that the nanosystem remained stable at low enzyme concentrations mimicking plasma conditions, whereas higher enzyme concentrations induced degradation of the albumin matrix and subsequent disaggregation of sAuNPs. Conclusions: Overall, this study presents a degradable, albumin-based sAuNP nanosystem with NIR-responsive properties and potential for nanomedicine applications to inhibit Aβ aggregation in AD. Full article
(This article belongs to the Topic Advanced Nanotechnology in Drug Delivery Systems)
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16 pages, 2451 KB  
Article
Physiological Stress Signatures of Waterborne Glyphosate Exposure in Apostichopus japonicus: Insights for Aquatic Ecotoxicology
by Jingchun Sun, Shaoping Kuang and Hongsheng Yang
Toxics 2026, 14(4), 282; https://doi.org/10.3390/toxics14040282 - 26 Mar 2026
Viewed by 540
Abstract
Glyphosate is a widely used herbicide with increasing concern regarding its non-target impacts in coastal ecosystems and mariculture species. Here, we profiled acute physiological stress signatures of waterborne glyphosate exposure in the sea cucumber Apostichopus japonicus, integrating measured exposure concentrations, tissue residues, [...] Read more.
Glyphosate is a widely used herbicide with increasing concern regarding its non-target impacts in coastal ecosystems and mariculture species. Here, we profiled acute physiological stress signatures of waterborne glyphosate exposure in the sea cucumber Apostichopus japonicus, integrating measured exposure concentrations, tissue residues, digestive and oxidative/innate immune biomarkers, and gut microbiota. After 24 h exposure, measured waterborne glyphosate confirmed the intended gradient (0.09 ± 0.02, 1.26 ± 0.09, and 4.49 ± 1.12 mg/L for low-, medium-, and high-dose treatments, respectively), and overt stress phenotypes with mortality occurred only at the high dose (36.67%), enabling separation of high-dose survivors (HS) and high-dose dead (HD) for downstream analyses. Tissue measurements showed low/background levels in controls, with compartment-specific distribution: the respiratory tree exhibited higher burdens at the medium dose, whereas coelomic fluid showed the highest burdens in HS at the 24 h endpoint. Functionally, most intestinal digestive enzymes were unchanged, but trypsin activity was consistently suppressed across exposed groups (p < 0.05). In coelomic fluid, oxidative stress responses were evident, with elevated MDA (L and M), reduced CAT (L, M, and HS), and reduced GSH-PX in HS (all p < 0.05), while SOD, GR, and lysozyme showed no significant changes. Gene sequencing of 16S rRNA (n = 3 per group) revealed significant shifts in community diversity/evenness (Shannon p = 0.0497; Simpson p = 0.0484) and beta diversity (PCo1 = 30.08%, PCo2 = 26.30%; PERMANOVA F = 1.816, p = 0.008), with LEfSe indicating discriminative taxa associated with exposure/outcomes. Collectively, these multi-level endpoints define an acute glyphosate stress signature in A. japonicus, linking internal dose distribution to oxidative disruption, impaired intestinal proteolysis, and microbiome restructuring. Full article
(This article belongs to the Section Ecotoxicology)
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15 pages, 266 KB  
Article
Preliminary Investigation on Salivary Enzymes of Massese Sheep
by Simona Sagona, Alessia Di Rosso, Francesca Coppola, Chiara Benedetta Boni, Claudia Russo, Lionella Palego, Laura Betti, Gino Giannaccini, Antonio Felicioli and Lucia Casini
Animals 2026, 16(7), 1008; https://doi.org/10.3390/ani16071008 - 25 Mar 2026
Viewed by 418
Abstract
Saliva is an important biological matrix that allows the investigation of various welfare parameters; in ruminants, it is abundant and can be easily collected without requiring professional veterinary intervention. The aim of the investigation was to provide additional information on both digestive and [...] Read more.
Saliva is an important biological matrix that allows the investigation of various welfare parameters; in ruminants, it is abundant and can be easily collected without requiring professional veterinary intervention. The aim of the investigation was to provide additional information on both digestive and antioxidant enzymes of sheep farmed in good welfare conditions, assessed with the Animal welfare indicators (AWIN) protocol. Small surgical forceps holding a Salivette® device (Sarstedt, Nümbrecht, Germany) were gently inserted into the sheep’s mouths without any force. The sheep chewed the swabs independently for a few seconds, allowing saliva collection. Seventeen enzymes from the saliva of 15 sheep were analyzed: antioxidant enzymes (catalase, glutathione S transferase), proteolytic enzymes (trypsin, chymotrypsin, N-aminopeptidases, carboxypeptidase A and B), carbohydrases (glucose oxidase, amylase, cellulase, lignin peroxidase, chitinase and α-glycosidase), and esterases (alkaline and acidic phosphatases, lipase and esterase). Esterase activity showed the highest value (12.95 ± 1.25 U/mg of proteins), whereas lignin peroxidase activity showed the lowest (2.23 ± 0.37 µU/mg of proteins). The activity of all enzymes was observed except for glutathione S transferase and α-glycosidase. Among the enzymes, lipase activity has already been identified as a biomarker of stress in sheep saliva. This investigation may represent a basis for further investigations into the diet and adaptive responses of sheep to different environmental conditions. Furthermore, samples collected using the Salivette® device can be easily obtained without requiring specialized staff and without causing any stress to the animals. Further investigations into the origin of individual enzymes using a proteomic approach are desirable. Full article
(This article belongs to the Section Small Ruminants)
24 pages, 7459 KB  
Article
The Impact of Lycium barbarum Polysaccharides on Growth Performance, Digestive Enzyme, Muscle and Skin Characteristics, and Immune-Antioxidant Functions in Coral Trout (Plectropomus leopardus)
by Chengkun Zhang, Chuanpeng Zhou, Zhengyi Fu and Zhenhua Ma
Fishes 2026, 11(3), 186; https://doi.org/10.3390/fishes11030186 - 20 Mar 2026
Viewed by 315
Abstract
This study investigated the effects of Lycium barbarum polysaccharides (LBP) supplementation on various indicators in coral trout (Plectropomus leopardus), including growth performance, digestive enzyme activity, muscle and skin morphology, inflammatory immune gene expression, as well as immune and antioxidant responses. In [...] Read more.
This study investigated the effects of Lycium barbarum polysaccharides (LBP) supplementation on various indicators in coral trout (Plectropomus leopardus), including growth performance, digestive enzyme activity, muscle and skin morphology, inflammatory immune gene expression, as well as immune and antioxidant responses. In the experiment, fish were fed diets supplemented with different concentrations of LBP (0%, 0.05%, 0.1%, 0.2%, 0.5%, and 1%) over a designated experimental period. The results showed that moderate supplementation of LBP significantly improved growth performance, with the optimal concentration being around 0.243%, achieving the highest specific growth rate. LBP supplementation also enhanced intestinal digestive enzyme activity, such as trypsin in the 0.1% and 1% groups, and α-amylase in the 0.5% group. Additionally, LBP improved the nutritional composition of muscle, with the 1% group showing higher crude protein content and the 0.2–1% groups having lower crude fat content. Moderate LBP supplementation improved skin color and pigmentation, increasing the brightness, redness, and yellowness of the dorsal skin, as well as boosting carotenoid and astaxanthin concentrations. It also enhanced the immune and antioxidant functions of the skin (e.g., SOD, CAT, GSH-Px, AKP, and LZ) and improved the immune functions of the mucus (e.g., C3, C4, IgM, IgT, AKP, and LZ). Furthermore, the expression of key pro-inflammatory genes, such as TNF-α and IL-1β, was reduced. These findings suggest that LBP can serve as a natural feed additive to enhance the overall quality and health of coral trout, contributing to sustainable aquaculture practices. Full article
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18 pages, 11105 KB  
Article
The Effects of Compound Chinese Herbal Medicine on the Growth and Digestive and Immune Systems of Megalobrama amblycephala
by Xijing Ye, Yunsheng Zhang, Hu Xia, Huangjie Fan, Jiahui Hu, Yanan Gong, Rurou Fu, Fuyan Chen and Liangguo Liu
Animals 2026, 16(6), 925; https://doi.org/10.3390/ani16060925 - 15 Mar 2026
Viewed by 477
Abstract
Chinese herbal medicine is rich in active ingredients that can promote growth and enhance immune function. In this study, Lycium barbarum, Panax ginseng, Astragalus membranaceus and Phragmitis rhizoma were crushed and mixed to prepare a compound Chinese herbal medicine. The basic [...] Read more.
Chinese herbal medicine is rich in active ingredients that can promote growth and enhance immune function. In this study, Lycium barbarum, Panax ginseng, Astragalus membranaceus and Phragmitis rhizoma were crushed and mixed to prepare a compound Chinese herbal medicine. The basic feed of Megalobrama amblycephala was supplemented with 0 (control group), 1% (T1), 2% (T2) and 4% (T3) of this compound medicine. After raising for 90 days, in the T1 and T2 experimental groups, the length and width of intestinal villi and the activities of amylase, trypsin and lipase in the intestine were significantly higher than those in the control group. The weight gain rate and specific growth rates were highest and the feed coefficient was lowest in the T2 experimental group. In the control group, a large number of dilated hepatic sinusoids were detected, while this number significantly decreased in the T1 experimental group and they were not detected at all in the T2 and T3 experimental groups. The spleen and liver body indices were highest in the T2 experimental group. In all experimental groups, the Lys content and the activities of T-SOD, CAT, ACP, AKP and GSH-PX in serum were significantly higher than those of the control group. The expression of IgM, C3, TNF-ɑ and IL-1β in the head kidney; C3, TNF-ɑ and IL-1β in the spleen; C3 and IL-1β in the gills; IgM, C3 and IL-1β in liver; and IL-1β in the intestine was highest in the T2 experimental group. After challenge with Aeromonas hydrophila, the cumulative mortality rate of M. amblycephala was lowest in the T2 experimental group. The results of this study indicated that this compound Chinese herbal medicine could significantly enhance immunity, increase the activity of intestinal digestion-related enzymes and promote the growth of M. amblycephala. The appropriate addition amount of this compound Chinese herbal medicine in the basic feed of M. amblycephala was 2%. Full article
(This article belongs to the Special Issue Advances in Fish Immunology: Novel Strategies for Disease Prevention)
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24 pages, 2277 KB  
Article
Optimized Solid-State Fermentation of Sugar Beet Pulp with Mixed Microbes Improves Its Nutritional Value and Promotes Growth, Health, and Intestinal Function in Yellow Catfish (Pelteobagrus fulvidraco)
by Ning Qiu, Tanqing Chi, Xuan Luo, Hao Yang, Chi Zhang, Hongsen Xu and Xin Liu
Animals 2026, 16(6), 915; https://doi.org/10.3390/ani16060915 - 14 Mar 2026
Viewed by 377
Abstract
The rising cost of conventional protein sources such as soybean meal has prompted the search for sustainable and economical alternatives in aquafeeds. Sugar beet pulp (SBP), an abundant by-product of the sugar industry, possesses nutritional potential but is limited by its high fiber [...] Read more.
The rising cost of conventional protein sources such as soybean meal has prompted the search for sustainable and economical alternatives in aquafeeds. Sugar beet pulp (SBP), an abundant by-product of the sugar industry, possesses nutritional potential but is limited by its high fiber and anti-nutritional factors. Solid-state fermentation (SSF) offers a promising approach to enhance its nutritive value and functional properties. This study evaluated the effects of dietary inclusion of mixed microbial solid-state fermented beet pulp (FBP) on the growth, systemic health and intestinal function of juvenile yellow catfish (Pelteobagrus fulvidraco). First, orthogonal optimization determined Lactiplantibacillus plantarum:Saccharomycopsis fibuligera:Bacillus subtilis = 1:3:3 as the optimal ratio, significantly improving the nutritional profile of FBP. Based on this optimized FBP, an 8-week feeding trial, five isonitrogenous and isolipidic diets were formulated by replacing 0–12% soybean meal with FBP. The results demonstrated that 9% FBP inclusion yielded optimal growth performance and significantly improved muscle texture. At the systemic level, FBP supplementation reduced serum lipid markers and liver enzyme activities while enhancing antioxidant capacity. At the intestinal level, FBP promoted intestinal health by increasing key digestive enzyme (lipase, trypsin, amylase) activities, stimulating villus development, and improving intestinal antioxidant status. Furthermore, gut microbiota analysis revealed that dietary FBP supplementation significantly modulated intestinal microbial composition, with notable enrichment of genera such as Leucobacter. In conclusion, FBP is a multi-functional ingredient that enhances growth, product quality, systemic physiology, and intestinal health in yellow catfish aquaculture. These findings provide a viable strategy for the sustainable utilization of agricultural by-products in aquafeeds. Full article
(This article belongs to the Special Issue Fish Nutrition, Physiology and Management: Second Edition)
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11 pages, 228 KB  
Article
A Pilot Proteomic Analysis of Tear Fluid in Domestic Cats with and Without Conjunctivitis Using MALDI–TOF/TOF Mass Spectrometry
by Takuya Yogo, Shotaro Iino and Kinya Katayama
Animals 2026, 16(6), 912; https://doi.org/10.3390/ani16060912 - 13 Mar 2026
Viewed by 351
Abstract
Feline conjunctivitis is a common ocular disorder; however, the molecular composition of feline tear fluid and its alterations during ocular surface inflammation remain poorly characterized. This pilot study aimed to explore the tear proteome of cats with conjunctivitis using matrix-assisted laser desorption/ionization time-of-flight [...] Read more.
Feline conjunctivitis is a common ocular disorder; however, the molecular composition of feline tear fluid and its alterations during ocular surface inflammation remain poorly characterized. This pilot study aimed to explore the tear proteome of cats with conjunctivitis using matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI–TOF/TOF MS) and to compare findings with those from clinically healthy cats. Tear samples were collected using Schirmer tear test strips from healthy cats and cats diagnosed with conjunctivitis. Total protein concentration was measured by ultraviolet absorbance spectrophotometry, and tear proteins were separated by SDS–polyacrylamide gel electrophoresis, followed by in-gel trypsin digestion and MALDI–TOF/TOF MS analysis. Nine distinct tear proteins were identified, including antimicrobial and immune-related components such as lactoperoxidase, lactotransferrin, albumin, and immunoglobulin A constant region. Lactoperoxidase and SBP1 were identified in feline tear fluid for the first time. No proteins uniquely associated with conjunctivitis were detected. The mean total tear protein concentration was numerically higher in cats with conjunctivitis (13.06 ± 0.75 mg/mL) than in healthy cats (9.69 ± 0.67 mg/mL); however, this difference did not reach statistical significance (p = 0.095) and should be interpreted cautiously given the limited sample size. This pilot study provides preliminary insights into tear protein profiles in cats with conjunctivitis and highlights the need for larger quantitative investigations. These findings provide a preliminary framework for future studies aimed at further characterizing molecular alterations associated with feline ocular surface disorders. Full article
24 pages, 8610 KB  
Article
Effects of Dietary Methionine and Lysine Balance on Intestinal Function and Immune Organ Development in Wanxi White Geese During the Brooding Period
by Shaoqi Shi, Changsheng Jiang, Suting Fang, Leilei Li, Xiaojin Li, Ahmed H. Ghonaim, Man Ren and Shenghe Li
Animals 2026, 16(6), 872; https://doi.org/10.3390/ani16060872 - 11 Mar 2026
Viewed by 439
Abstract
(1) Background: This experiment investigated the effects of balancing methionine or lysine under different dietary protein levels on intestinal function and immune organ development in Wanxi white geese during the brooding period. (2) Methods: A total of 180 one-day-old Wanxi white geese were [...] Read more.
(1) Background: This experiment investigated the effects of balancing methionine or lysine under different dietary protein levels on intestinal function and immune organ development in Wanxi white geese during the brooding period. (2) Methods: A total of 180 one-day-old Wanxi white geese were randomly divided into six groups using a 2 × 3 factorial design, with amino acid balance (lysine or methionine) and dietary crude protein levels (20%, 18%, and 16%) as experimental factors. Each group consisted of five replicates with six geese per replicate. The feeding trial lasted 28 days. (3) Results: The lysine-balanced diet significantly increased jejunal lipase activity in 14-day-old geese (p < 0.05). In contrast, the methionine-balanced diet significantly enhanced ileal trypsin, maltase, and amylase activities at 14 days, as well as ileal trypsin activity at 28 days (p < 0.05). Lysine balance markedly improved jejunal weight and muscularis thickness, ileal weight, villus height, and villus-to-crypt (V/C) ratio in 14-day-old geese, and further increased jejunal villus height and ileal crypt depth and muscularis thickness at 28 days (p < 0.05). Methionine balance significantly elevated total antioxidant capacity (T-AOC) in the jejunum and ileum of geese at both 14 and 28 days (p < 0.05). Moreover, the methionine-balanced group exhibited a significantly higher thymus index and spleen index in 28-day-old geese compared with the lysine-balanced group (p < 0.05). Notably, geese fed the 18% crude protein diet showed significantly greater follicular and medullary areas of the bursa of Fabricius at 14 days than those fed 20% or 16% protein diets (p < 0.05). (4) Conclusions: Appropriate dietary protein reduction combined with balanced lysine or methionine supplementation effectively enhances intestinal digestive function, antioxidant capacity, and immune organ development in Wanxi white geese. Among the tested treatments, a dietary protein level of 18% produced the most favorable overall outcomes during the brooding period. Full article
(This article belongs to the Special Issue Optimizing Alternative Protein Sources for Sustainable Poultry Diet)
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18 pages, 2658 KB  
Article
Abridged Ribosome Profiling for Accurate Bacterial Translation Measurements
by Marc Follmer, Korbinian Pürckhauer and Klaus Neuhaus
Methods Protoc. 2026, 9(2), 45; https://doi.org/10.3390/mps9020045 - 10 Mar 2026
Viewed by 630
Abstract
Ribosome profiling, or Ribo-Seq, is a powerful tool for studying translation. It maps the positions of translating ribosomes on mRNAs, providing insights into actively expressed genes. Unlike mass spectrometry, Ribo-Seq is not affected by the same biases that limit mass spectrometry, such as [...] Read more.
Ribosome profiling, or Ribo-Seq, is a powerful tool for studying translation. It maps the positions of translating ribosomes on mRNAs, providing insights into actively expressed genes. Unlike mass spectrometry, Ribo-Seq is not affected by the same biases that limit mass spectrometry, such as protein size, concentration, trypsin digestibility, or hydrophobicity. Thus, the translatome has previously been used to discover unannotated genes, including small and overlapping ones that were missed by mass spectrometry or gene prediction models. However, a major limitation of classical ribosome profiling is its complexity, involving multiple steps such as sucrose density gradient centrifugation and gel electrophoresis. These make the method costly, time-consuming, and limit its throughput. Here, we compared the classical method using gradient centrifugation and size exclusion by gel electrophoresis with shortened versions to evaluate experimental performance and achieved reductions. Our results show that the sucrose density gradient centrifugation is essential for obtaining accurate Ribo-Seq data, whereas gel electrophoresis for size selection can be omitted (although this requires increased sequencing depth). Thus, future experiments can be conducted with reduced sample input and hands-on time while still achieving a reliable quantification of translation. Full article
(This article belongs to the Section Molecular and Cellular Biology)
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16 pages, 2683 KB  
Article
Entomopathogenic Effects of the Plant-Associated Fungus Ochroconis guangxiensis X22 Strain on the Physiological and Metabolic State of the Rice-Pest Planthopper, Sogatella furcifera
by Yanxin Yu, Fenghua Zeng, Yanyan Long, Zhengxiang Sun, Xinghao Wang, Bixia Qin, Jihui Yu, Wenlong Zhang, Yan Zhang and Ling Xie
Agriculture 2026, 16(5), 567; https://doi.org/10.3390/agriculture16050567 - 2 Mar 2026
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Abstract
The white-backed planthopper (Sogatella furcifera) is a major pest in rice-growing regions worldwide. It severely limits rice production through piercing–sucking feeding, oviposition injury, and by efficiently transmitting the Southern Rice Black-Streaked Dwarf Virus (SRBSDV). Previous studies demonstrated that the dark septate [...] Read more.
The white-backed planthopper (Sogatella furcifera) is a major pest in rice-growing regions worldwide. It severely limits rice production through piercing–sucking feeding, oviposition injury, and by efficiently transmitting the Southern Rice Black-Streaked Dwarf Virus (SRBSDV). Previous studies demonstrated that the dark septate endophytic fungus Ochroconis guangxiensis strain X22 exhibits control activity against SRBSDV. To further evaluate its biocontrol potential, this study investigated the effects of the X22 strain on S. furcifera, the primary vector of SRBSDV. In this study, we established an X22–rice symbiotic system to evaluate its effects on the biological traits of S. furcifera. The results showed that, compared with a clear water treatment, the X22 strain significantly reduced the feeding amount (29.02%), egg-laying amount (12.30%), and hatching rate (11.58%) of S. furcifera. Gene expression analysis showed that the relative expression levels of the Target of Rapamycin (TOR) and vitellogenin (Vg) genes in one-day-old S. furcifera from the X22 treatment group were modestly downregulated, although no significant differences were detected compared with the control. Enzyme activity assays revealed that between 72 and 120 h post-treatment, the activities of detoxification enzymes, including carboxylesterase (CarE) and acetylcholinesterase (AChE), generally declined following X22 exposure. In contrast, the activities of protective enzymes, superoxide dismutase (SOD) and catalase (CAT), as well as certain digestive enzymes, α-amylase (α-AL) and trypsin, were induced. Conversely the activities of glutathione peroxidase (GSH-Px) and lipase (LPS) were suppressed. However, the physiological mechanisms underlying its effect on S. furcifera remain unclear. Collectively, these results demonstrate that the O. guangxiensis X22 strain inhibits S. furcifera reproduction by disrupting its physiological metabolism through multiple pathways, providing a mechanistic basis for its development as an environmentally friendly biocontrol agent. Full article
(This article belongs to the Section Crop Protection, Diseases, Pests and Weeds)
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Article
Survival and Metabolic Enzyme Response of Spodoptera exigua Larvae Under Different Nutritional Conditions
by Hongzhi Zhang, Xin Gao, Fengjiao Qiao, Ziyu Shao, Yan Shi, Bin Zhang and Chuande Zhao
Agronomy 2026, 16(4), 415; https://doi.org/10.3390/agronomy16040415 - 9 Feb 2026
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Abstract
Spodoptera exigua has emerged as a globally agricultural pest due to its strong adaptability to diverse nutritional conditions. In this study, we used artificial diets with a protein-to-carbohydrate (P:C) ratio of 1:1 as the control and examined the effects of extreme nutritional imbalances [...] Read more.
Spodoptera exigua has emerged as a globally agricultural pest due to its strong adaptability to diverse nutritional conditions. In this study, we used artificial diets with a protein-to-carbohydrate (P:C) ratio of 1:1 as the control and examined the effects of extreme nutritional imbalances (P:C = 1:7 or 7:1) on survival and the activities of nine enzymes in third-stage larvae. The results showed that survival rates of larvae in both unbalanced diet groups were lower than those in the control group. High-carbohydrate-low-protein diets enhanced carboxylesterase, polyphenol oxidase, superoxide dismutase, peroxidase and three digestive enzymes (trypsin, amylase, lipase), cope with overeating and peroxide accumulation. The high-protein-low-carbohydrate diet exclusively increased lipase activity, confirming that larvae compensate for carbohydrate deficiency through lipid mobilization. These findings provide novel insights into polyphagy mechanisms in S. exigua, establishing a theoretical basis for predicting pest dispersal and developing control strategies. Full article
(This article belongs to the Section Pest and Disease Management)
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