β-Conglycinin (7S), a major soybean allergen traditionally considered susceptible to degradation during gastrointestinal digestion, still induces diarrhea and intestinal inflammation in young animals and susceptible individuals, suggesting that its biological fate and pro-inflammatory activity are more complex than previously recognized. Our preliminary study demonstrated that 7S could self-assemble into digestion-resistant β-conglycinin nanoparticles (7SNs) in the small intestine of weaned piglets. This study was designed to elucidate the intracellular trafficking behavior and inflammatory mechanisms of 7SNs in intestinal porcine epithelial cells (IPEC-J2).The results showed that 7SNs were internalized via clathrin- and caveolae-mediated endocytosis and subsequently accumulated in lysosomes. After lysosomal localization, 7SNs activated the lysosomal Ca2+ channel transient receptor potential mucolipin 1 (TRPML1), thereby inducing lysosomal Ca2+ release and promoting transcription factor EB (TFEB) nuclear translocation and activation, ultimately triggering lysosomal exocytosis. Inhibition of TFEB activation, lysosomal Ca2+ release, or Ca2+-dependent lysosomal exocytosis markedly suppressed both 7SNs exocytosis and the secretion of pro-inflammatory cytokines, including IL-1β and TNF-α. These findings demonstrated that 7SNs induced inflammatory responses in IPEC-J2 cells through the lysosomal exocytosis pathway mediated by TRPML1/Ca2+/TFEB. Collectively, this study demonstrates that the in vivo nanoscale remodeling of dietary proteins drives intestinal inflammation via enterocytic endocytosis and lysosomal exocytosis, providing new insights into food protein-related intestinal inflammation.

High-yielding dairy cows often experience metabolic stress during early lactation, leading to subclinical ketosis (SCK) and reproductive impairment. This study investigates how SCK-associated conditions (hypoglycemia and elevated nonesterified fatty acids, NEFA) affect bovine granulosa cell (GC) function via the PI3K/AKT pathway. Primary GCs were cultured under four metabolic conditions for 24 h: normal glucose (NG), low glucose (LG), NG + high NEFA, and LG + high NEFA. Additional groups treated with the PI3K/AKT activator SC79 or inhibitor LY294002 under LG + NEFA stress were included. Combined LG + NEFA stress reduced cell viability, increased apoptosis, impaired steroidogenesis, and disrupted mitochondrial function. While LG alone increased p-AKT, NEFA alone suppressed this. SC79 rescued these effects, whereas LY294002 exacerbated them. These findings demonstrate that metabolic stressors disrupt GC proliferation, steroidogenesis, and mitochondrial homeostasis through PI3K/AKT pathway dysregulation, offering insights into the SCK-associated reproductive dysfunction.

BACKGROUND:Feline hepatic lipidosis (FHL) lacks well-defined metabolic biomarkers and mechanistic understanding. HYPOTHESIS/OBJECTIVES:Clarify the metabolic biomarkers and mechanisms of FHL. ANIMALS:Two groups of cats were analyzed: 14 cats (7 FHL cases and 7 healthy controls) for liver metabolomics, and 60 cats (10 FHL cases and 50 healthy controls) for blood 3-hydroxybutyrate (3-HB) testing. METHODS:Hepatic lipidosis was confirmed by histopathological assessment and ultrasonography. Untargeted metabolomics was performed using gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS), with pathway analysis using MetaboAnalyst 5.0. Serum 3-HB was quantified using an enzymatic assay. Statistical analysis included principal component analysis, orthogonal partial least squares discriminant analysis, cross-validation, permutation tests, and F₁ score evaluation for model validation. RESULTS:Gas chromatography-mass spectrometry identified four metabolites with variable importance in projection > 1 (bootstrap 95% stability range, 0.9-1.3) and < 0.05, whereas LC-MS identified 243 differentially abundant metabolites (101 upregulated, 142 downregulated), with 19 demonstrating diagnostic potential (area under the curve [AUC] > 0.7). Pathway analysis identified perturbations in vitamin B6 metabolism (P = .003) and fructose/mannose metabolism (P < .001). Serum 3-HB underwent analytical validation in 60 specimens (10 FHL cases, 50 healthy controls), which identified concordant hepatic and systemic increases in FHL. At the optimal cutoff (>2.43 mmol/L), it achieved an AUC of 0.86 (95% confidence interval [CI], 0.65-1.00) with 92% sensitivity (95% CI, 89%-100%) and 88% specificity (95% CI, 74%-100%) in distinguishing FHL from normal liver under controlled laboratory conditions. CONCLUSIONS AND CLINICAL IMPORTANCE:Preliminary data suggest FHL involves dysregulated fatty acid and vitamin B6 metabolism. Although serum 3-HB shows promise, future studies should include disease controls (eg, cholangitis, hepatitis) to establish specificity. These findings provide foundational insights for mechanistic research.

During clinical and subclinical Escherichia coli mastitis, bovine mammary epithelial cells (BMEC) are stimulated by LPS, leading to cellular inflammatory response, oxidative stress, and autophagy. This study hypothesized that rutin might mitigate these damages in BMEC via modulation of specific signaling pathways. To simulate mammary gland inflammation, different concentrations of LPS were used to stimulate BMEC. Different concentrations of rutin were used to pretreat the BMEC. The results demonstrated that LPS stimulated the NLR family pyrin domain containing 3 (NLRP3) inflammasome and its downstream effector molecules to produce an inflammatory response. Lipopolysaccharide caused a decrease in antioxidant enzymes and the total antioxidant capacity, an increase in malondialdehyde (MDA) levels, and a decrease in silent mating type information regulation 2 homolog-1 (SIRT1) levels. Treatment with LPS altered the morphological structure of the cells, converted the autophagy marker microtubule-associated protein light chain 3 (LC3)-I into LC3-II, and the increased the level of Beclin-1. Rutin increased silent mating type information regulation 2 homolog-1 (SIRT1) activity, reduced reactive oxygen species (ROS) generation, and decreased specific oxidative stress markers MDA. Additionally, rutin enhanced antioxidant enzyme activities including total antioxidant capacity, superoxide dismutase, catalase, and glutathione peroxidase. Rutin also inhibited NLRP3 inflammasome activation, which is associated with ROS production and oxidative stress. The mechanism of the protective effect of rutin on BMEC was investigated using the SIRT1 specific inhibitor EX-527. EX-527 considerably weakened the regulating function of rutin and aggravated the LPS-induced damage to BMEC. Rutin inhibited NLRP3 inflammasome activation and the downstream effector molecule gasdermin D (GSDMD) through SIRT1, thus alleviating excessive autophagy and reducing the inflammatory damage of cells. This study confirms that rutin reduces LPS-induced inflammation, autophagy, and oxidative stress in BMEC through the SIRT1/NLRP3 pathway. These findings provide new insights into addressing mastitis in dairy cows and offer promising clinical applications for future therapeutic interventions.

Zearalenone (ZEA) is a mycotoxin commonly found in moldy cereals and has a range of toxic effects that have seriously affected animal husbandry. Rutin, a natural flavonoid with antioxidant activities, has been studied for its potential involvement in mitigating ZEA-induced apoptosis in porcine endometrial stromal cells (ESCs) and its potential molecular mechanism, particularly concerning the expression of Nrf2. This study investigates the molecular pathways by which rutin alleviates ZEA-induced ESC apoptosis, focusing on the role of Nrf2. Experimental data reveal that ZEA suppresses Nrf2 nuclear translocation and reduces mitochondrial membrane potential (MMP), leading to oxidative stress, endoplasmic reticulum stress (ERS), and mitochondrial pathway-driven apoptosis. Notably, rutin mitigates ZEA-induced apoptosis through Nrf2 activation. These findings highlight Nrf2 as a critical factor in rutin’s protective effects against ZEA-induced apoptosis, offering valuable insights for the clinical prevention and treatment of ZEA toxicity.

Transition dairy cows face severe oxidative stress that disrupts mammary epithelial homeostasis through intertwined oxidative, inflammatory, and endoplasmic reticulum (ER) stress pathways. This study hypothesized that rutin, a natural flavonoid, alleviates hydrogen peroxide (H2O2)-induced oxidative damage in bovine mammary epithelial cells (BMECs) via AMPK/NFE2L2 signaling activation. In this study, BMECs were pre-incubated with rutin. Subsequently, cells were treated with or without H2O2. Additionally, by transfecting BMECs with NFE2L2 siRNA (siNFE2L2), we investigated how AMPK/NFE2L2 signaling mediated by rutin may prevent H2O2-induced oxidative damage. The results show that increases in reactive oxygen species (ROS), expression of inflammatory cytokines, expression of proteins related to endoplasmic reticulum stress and the apoptosis rate induced by H2O2 in cells, were attenuated in rutin cultures. Challenges with H2O2 led to a lower abundance of proteins related to AMPK and NFE2L2. Comparatively, these effects were reversed in cultures with rutin. Transfection with siNFE2L2 reversed the protection of rutin, suggesting that NFE2L2 is essential for the protective mechanism of rutin. These results elucidated the molecular mechanism of rutin’s resistance to H2O2-mediated oxidative injury through the AMPK/NFE2L2 signaling pathway and suggested that it could be used as a potent in vivo antioxidant for ruminants during periods of stress, such as before and after calving.

SIRT1-SREBP−1c/PGC−1α signaling is involved in the production of non-esterified fatty acids (NEFAs) and liver lipid metabolism disorders in ketotic calf. The molecules contained in extracellular vesicles (EVs) regulate intercellular communication, and research on calf hepatocytes−derived EVs has become a hot spot. We hypothesized that EVs in cell culture supernatants could affect lipid metabolism in hepatocyte models via SIRT1/SREBP−1c/PGC−1α signaling. Non-ketosis (NK, 0 mM NEFA) and clinical ketosis calf models (CK, 2.4 mM NEFAs) were established in vitro cultured calf hepatocytes and EVs were extracted from their supernatants as NK−derived EVs and CK−derived EVs, respectively. In vitro hepatocyte models, comprising a normal culture group (normal) and the group treated with NEFAs at 2.4 mM (2.4 NEFA), were treated with NK and CK−derived EVs. In addition, we transfected an SIRT1−overexpressing adenovirus into calf hepatocytes and determined the expression of key genes, enzymes, and proteins involved in the SIRT1/SREBP−1c/PGC−1α pathway. The results showed that the NK−derived EVs inhibited the expression of the SREBP−1c gene and protein and increased the expression of the SIRT1 and PGC−1α genes and proteins (p < 0.05). In contrast, CK−derived EVs induced lipid metabolism disorders in the normal hepatocyte group and aggravated NEFA-induced lipid metabolism imbalances in hepatocytes (p < 0.05). Moreover, overexpression of SIRT1 confirmed that EVs exert vital functions in hepatocyte lipid metabolism via SIRT1/SREBP−1c/PGC−1α signaling to regulate hepatocyte lipid metabolism. In summary, NK−derived EVs alleviated liver lipid metabolism disorders caused by NEFAs via modulation of SIRT1/SREBP−1c/PGC−1α signaling, while CK−derived EVs had the opposite effect. NK−derived EVs upregulated lipid oxidation-related genes and downregulated lipid synthesis-related genes, suggesting that NK−derived EVs could be used as biological extracts to alleviate lipid metabolism disorders in ketotic calf.

Feline hepatic lipidosis (FHL) is a common liver dysfunction caused by metabolic disorders. The objective was to evaluate the metabolic alteration in the cats of FHL and to identify biomarkers that can serve as biomarker for FHL. Differential metabolites in the serum of spontaneous FHL cats (FS, n = 12) and healthy cats (CS group, n = 12) were analyzed using GC/MS metabolomics. Differential metabolites with diagnostic significance were identified through receiver operating characteristic (ROC) curves. The expression level of the differential metabolite 2-hydroxybutyric acid (2-HB) was detected in the serum of the FS and CS groups, and biomarker were established. The biomarker efficacy of 2-HB for FHL was verified using serum samples from cats with FHL caused by different etiologies (F, n = 10) and healthy cats (C, n = 50). There were 13 significantly different metabolites between the CS and FS groups (VIP > 1, P < 0.05) with the area under the ROC curve (AUC) greater than 0.70. The AUC for serum 2-HB was 0.90 (95% confidence interval 0.767-1.000, P < 0.001), with an optimal critical value of 564.8 ng/L. By randomly detecting serum 2-HB in groups F and C (the optimal cut-off value is 564.8 ng/L), the detection rate for FHL diagnosis was 100% and the false positive rate was 0%. In cats with FHL, metabolic changes occur in amino acids, nucleotide sugars, glycerophospholipids, phenylalanine, galactose, alpha-linolenic acid, and glycerides. A serum 2-HB level greater than 564.8 ng/L serves as a biomarker for FHL.

Zearalenone (ZEA) is commonly found in crops and feed, with a high detection rate and concentration. This substance adversely affects reproduction and development, with particularly noticeable effects on pigs. Given these detrimental effects, there is an urgent need to identify effective protective agents against ZEA toxicity. Sulforaphane (SFN) has emerged as a promising candidate owing to its widely acknowledged antioxidant, anti-inflammatory, and detoxifying properties. This study elucidates ZEA's mechanism of oxidative injury in porcine endometrial stromal cells (ESCs) via mitochondrial-associated membrane (MAM) disruption and demonstrates SFN's protective role. ZEA disrupts the structural integrity and functional dynamics of mitochondrial-associated membranes (MAM) by downregulating PACS 2, leading to elevated intracellular Ca2+ levels (P < 0.01), increased ROS generation (P < 0.01), MDA accumulation (P < 0.01), and suppressed antioxidant enzyme activity (P < 0.01). SFN (10 μM) or the overexpression of PACS 2 can reduce the toxic damage caused by ZEA (45 μM). This study highlights the mechanism by which ZEA causes oxidative damage in porcine cells through mitochondrial membrane disruption and showcases SFN's protective effects, opening up possibilities for broader applications against ZEA toxicity.

Soybean glycinin (11S) and β-conglycinin (7S) are major contributors to allergic diarrhea and intestinal barrier damage in young animals. This study investigated the molecular mechanisms underlying the 7S- and 11S-induced dysfunction of mitochondrial and endoplasmic reticulum (ER) interactions in porcine intestinal epithelial (IPEC-J2) cells via the oxidative stress pathway. The results showed that 7S- and 11S-induced oxidative stress, as evidenced by the following findings: reduced manganese superoxide dismutase (Mn-SOD) activity and elevated 8-Hydroxy-2'-deoxyguanosine (8-OHdG) levels, with excessive reactive oxygen species (ROS) accumulation and elevated Ca2+ levels; decreased mitochondrial membrane potential (MMP), damaged mitochondria-associated endoplasmic reticulum membranes (MAM) structure; up-regulated the protein expression of glucose-regulated protein 75 (GRP75) and mitochondrial Rho-GTPase 1 (Miro1), while inositol 1,4,5 -trisphosphate receptor (IP3R), voltage-dependent anion channel 1 (VDAC1), mitofusin2 (MFN2) and phosphofurin acidic cluster sorting protein 2 (PACS2) were down-regulated. N-acetylcysteine (NAC)pre-treatment alleviated ROS accumulation and mitigated Ca2+ overload and MAM dysfunction, thereby ameliorating IPEC-J2 cell injury. In conclusion, 7S- and 11S-induced ROS burst to disrupt mitochondria-ER interaction homeostasis, leading to MAM structural damage and calcium dysregulation in IPEC-J2 cells, NAC effectively mitigated this process by scavenging ROS. These findings elucidate the critical involvement of subcellular organelle interaction disorders in food allergy pathogenesis and provide novel insights for targeted intervention strategies.

Zearalenone (ZEA) is an environmentally widespread mycotoxin capable of posing a serious threat to food safety and public health, and porcine endometrial stromal cells (ESCs) are particularly sensitive to the toxic effects of ZEA. We hypothesized that Rutin, a flavonoid antioxidant, could significantly alleviate ZEA-induced ferroptosis through the p53 signaling pathway. In this study, we used porcine ESCs as a research model. When porcine ESCs were co-cultured with the addition of Rutin and ZEA following p53 gene silencing via siRNA transfection, Rutin significantly mitigated ZEA-induced mitochondrial damage, oxidative stress, and Fe2 + content through the p53 pathway. Additionally Rutin lowered the expression of p53, ALOX12, and ACSL4 while significantly improving cytokinesis, antioxidant enzyme activity, and SLC7A11, GPX4, Nrf2, FTH1, thereby inhibiting cellular ferroptosis. These findings suggested a novel programmed death mechanism for alleviating the cytotoxic effects of ZEA, involving the knockdown of p53.

Deoxynivalenol (DON) and copper (Cu) are crucial food-related contaminants associated with health risks in both animals and humans. This study investigated the individual and combined effects of DON and Cu on a mouse hepatocyte line NCTC1469 cells. The results demonstrated that both DON and Cu induced ultrastructural damage, promoted mitochondrial vacuolization, increased pro-inflammatory cytokine levels, such as interleukin-1β (IL-1β) and tumor necrosis factor-alpha, reduced cell viability, and suppressed antioxidant enzyme activity, including glutathione and superoxide dismutase. Notably, DON + Cu enhanced cell viability compared to DON alone. In addition, co-treatment significantly reduced IL-1β levels relative to DON alone at 0.625, 1.25, and 2.5 μM. Transcriptome sequencing revealed that both DON alone and DON-Cu co-exposure triggered numerous differentially expressed genes, which were notably enriched in autophagy related pathways, such as ribosome biogenesis in eukaryotes, lysosome, spliceosome, and cell cycle. Meanwhile, the relative protein ratio of LC3-II/LC3-I was elevated at 0.65 μM DON, while the p62 expression was decreased in a dose-dependent manner compared to the control. In summary, DON exerts toxic effects on mouse hepatocytes, while Cu can mitigate DON-induced cellular damage at low concentrations, likely through involvement in Beclin-1/p62 related autophagic regulation.

High levels of non-esterified fatty acids (NEFA) in cows with subclinical ketosis (SCK) impair postpartum follicular development and disrupt estrus. The precise mechanism through which NEFA impacts the functionality of bovine follicular cells remains elusive. An in vivo experiment was conducted to compare SCK cows without estrus (SCK-E, n = 6) with healthy cows in estrus (C-E, n = 6). In the vitro test, bovine granulosa cells (GCs) were exposed to 0.4 mM NEFA. Notably, the SCK-E group exhibited an elevated ratio of phosphorylated adenosine 5'-monophosphate-activated protein kinase α (AMPKα) to total AMPKα in both liver and ovarian tissues, compared to the C-E group. NEFA treatment of GCs adversely affected steroid hormone synthesis, suppressed the expression of cyclin and proteins crucial for steroid synthesis, and triggered cell apoptosis, thereby inhibiting cell proliferation. Furthermore, it led to a decline in cell mitochondrial membrane potential and an increase in reactive oxygen species production, ultimately causing cellular damage. Subsequently, GCs were co-cultured with adenovirus (ad-AMPKα-siRNA) and NEFA (0.4 mM). Inhibiting AMPKα further exacerbated the detrimental effects of NEFA on steroid hormone synthesis, cell apoptosis, cell proliferation, and mitochondrial function in GCs. Furthermore, upon inhibiting AMPKα, a reduction was observed in both mRNA and protein levels of acetyl-CoA carboxylase 1, accompanied by an elevation in the levels of carnitine palmitoyltransferase-1. These findings suggest that AMPKα becomes activated in SCK cows experiencing elevated NEFA levels, and that AMPKα has the potential to mitigate the detrimental effects of NEFA on GCs function in vitro.

In dairy cows, the occurrence of subclinical ketosis (SCK) is particularly high during early lactation. Previously, we documented alterations in the abundance of adiponectin (ADPN) in anestrus cows with SCK in comparison to cows in estrus. In the present study, 60 cows were divided into two groups: control (C, n = 30) and SCK (n = 30). Based on cow's estrus situation in two group at 55-60 days postpartum, 15 anestrus SCK cows and estrus cows were designated the SCK-A group and C-E group, respectively. The SCK-A group had downregulated serum and follicular fluid ADPN levels compared with the C-E group. The serum ADPN level was positively correlated with the insulin level and follicle growth rate, and there was a positive correlation between ADPN and glucose in the follicular fluid. Primary culture of dairy cow granulosa cells (GCs) was established to observe the effect of low glucose (Glu) and/or ADPN on GCs cyclins and proteins important for steroid synthesis. The results showed that the addition of 1 µg/mL ADPN alleviated the negative effects of low Glu treatment on the proliferation of GCs and the expression of steroid secretion related protein proteins. Treatment with LY294002 (PI3K inhibitor) four experimental GCs groups: control (0 µg/mL ADPN), 1 µg/mL ADPN, LY294002 inhibitor, and 1 µg/mL ADPN+LY294002. The results showed that ADPN promotes the secretion of steroid hormones by GCs through the PI3K-AKT. In summary, ADPN plays a crucial role in ameliorating postpartum anestrus in dairy cows with SCK.

It is very common that high-producing dairy cows and the transition dairy cows experience oxidative stress. Excessive oxidative stress may cause inflammatory, apoptosis and eddoplasmic reticulum stress of bovine mammary epithelial cells (BMEC). Rutin, a natural flavonoid compound, has revealed remarkable antioxidative activity. The objective of this study was to evaluate effect of rutin on oxidative damage, including oxidaive stress, inflammatory, apoptosis and eddoplasmic reticulum stress, induced by H2O2 in BMEC and explore the corresponding mechanisms. In this study, BMEC were pre-incubated with rutin. Subsequently, cells were treated with or without H2O2. Additionally, to determine the role of AMPK/NEF2L2 signaling pathway in ruitn protected against H2O2-induced oxidative damage in BMEC, the cells were transfected with NFE2L2 siRNA (siNFE2L2). The results shown that, the increases of intracellular reactive oxygen species (ROS), inflammatory cytokines (TNF-α, IL-6, and IL-1β) expression, eddoplasmic reticulum stress related protein expression (GRP78 and CHOP) and the apoptosis rate induced by H2O2 were attenuated in the rutin cultures. The challenge with H2O2 led to lower abundance of proteins related to AMPK and NFE2L2. While these effects were reversed in cultures with rutin. The transfection with siNFE2L2 revoked the protection of rutin, suggested that NEF2L2 is essential for the protective mechanism of rutin. These results not only illuminate the molecular mechanism of AMPK/NFE2L2 signaling pathway involved in the protection of rutin against H2O2-induced oxidative damage, but also provide a potent antioxidant in vivo that could be administered to ruminant animals during stressful periods such as around the time of calving.

Aflatoxin B 1 (AFB 1 ) is known to inhibit growth, and inflict hepatic damage by interfering with protein synthesis. Allicin, has been acknowledged as an efficacious antioxidant capable of shielding the liver from oxidative harm. This study aimed to examine the damage caused by AFB 1 on bovine hepatic cells and the protective role of allicin against AFB 1 -induced cytotoxicity. In this study, cells were pretreated with allicin before the addition of AFB 1 for co-cultivation. Our findings indicate that AFB 1 compromises cellular integrity, suppresses the expression of nuclear factor erythroid 2 -related factor 2 (Nrf2). In addition, allicin attenuates oxidative damage to bovine hepatic cells caused by AFB 1 by promoting the expression of the Nrf2 pathway and reducing cell apoptosis. In conclusion, the results of this study will help advance clinical research and applications, providing new options and directions for the prevention and treatment of liver diseases.

Aflatoxin B1 (AFB1) is commonly found in feed ingredients and foods all over the world, posing a significant threat to food safety and public health in animals and humans. Lactobacillus salivarius (L. salivarius) was recorded to improve the intestinal health and performance of chickens. However, whether L. salivarius can alleviate AFB1-induced hepatotoxicity in geese was unknown. A total of 300 Lande geese were randomly assigned to five groups: control group, AFB1 low-dose group (L), L. salivarius+AFB1 low-dose group (LL), AFB1 high dosage groups (H), L. salivarius+AFB1 high dosage groups (LH), respectively. The results showed that the concentrations of ALT, AST, and GGT significantly increased after exposure to AFB1. Similarly, severe damage of hepatic morphology was observed including the hepatic structure injury and inflammatory cell infiltration. The oxidative stress was evidenced by the elevated concentrations of MDA, and decreased activities of GSH-Px, GSH and SOD. The observation of immunofluorescence, real-time PCR, and western blotting showed that the expression of PINK1 and the value of LC3II/LC3I were increased, but that of p62 significantly decreased after AFB1 exposure. Moreover, the supplementation of L. salivarius effectively improved the geese performance, ameliorated AFB1-induced oxidative stress, inhibited mitochondrial mitophagy and enhanced the liver restoration to normal level. The present study demonstrated that L. salivarius ameliorated AFB1-induced the hepatotoxicity by decreasing the oxidative stress, and regulating the expression of PINK1/Parkin-mediated mitophagy in the mitochondria of the geese liver. Furthermore, this investigation suggested that L. salivarius might serve as a novel and safe additive for preventing AFB1 contamination in poultry feed.