Search Results (215)

Essential oils (EOs) have gained recognition as promising alternatives to antibiotics due to their positive effects on bird growth performance, enhanced meat quality, and improved overall health, without producing the negative consequences associated with antibiotics. This study evaluated the effects of dietary supplementation of tea tree (TTEO) and thyme (TEO) EOs, individually or in combination with the probiotic BioPlus 2b (Bacillus subtilis and Bacillus licheniformis), on poultry broiler performance, including the meat quality. A total of 240 ROSS 308 broilers were assigned to eight dietary treatments over a 35-day trial. Parameters such as body weight (BW), feed conversion ratio (FCR), carcass portion, drip loss, and meat pH were evaluated. TTEO had a significant (p ≤ 0.05) impact on final carcass and breast portion, while in combination with probiotics, specifically TTEO with BioPlus significantly (p ≤ 0.05) reduced meat drip loss. GC-MS analysis identified terpinen-4-ol and γ-terpinene as the major constituents of TTEO, and thymol and carvacrol as the major constituents of TEO. In conclusion, combinations of TTEO, TEO, and probiotics can have beneficial effects on chicken raw meat quality, providing a complementary benefit to the industry and representing a viable alternative to conventional agents. Full article

The growing consumption of oilseed-pressed cakes (OSCs), a largely underutilized feedstock, plays a significant role in animal feed. The study evaluates the use of three OSCs—flax (FSC), pumpkin (PSC), and hemp (HSC)—as substrates for Bacillus subtilis ATCC 6051a (BS) in a solid-state fermentation (SSF) to enhance enzyme production and probiotic viability. The SSF process was assessed to evaluate the microbial growth, sporulation efficiency, enzymatic activity (protease, cellulase, xylanase, and phytase), and in vitro digestibility of fermented substrates. The results indicate that bacterial growth and sporulation varied significantly among substrates (p < 0.05). FSC presents the highest spore resistance (86.52%), followed by PSC (82.87%) and HSC (81.29%). Notably, protease was highest in HSC (184.67 U/g), while FSC supported maximum cellulase activity. HSC exhibited superior xylanase (1.86 ± 0.043 U/g DW, p < 0.05) and phytase production, while pH analysis indicated a shift toward alkalinity in PSC and HSC due to proteolytic activity. FSC maintained the most stable bacterial population during digestion, suggesting its potential as a probiotic carrier. These findings highlight that fermentation of OSCs with BS improved their nutritional value and can be used as a sustainable solution in feeding programs for piglets. Full article

Tiliacora triandra (Yanang) leaf contains polyphenols, flavonoids, and mucilage polysaccharides with antioxidant and prebiotic functions, making it a promising substrate for probiotic fermentation. This study aimed to optimize Yanang extraction and sterilization to preserve bioactive mucilage and support probiotic survivability during freeze-drying–based encapsulation, and evaluate antimicrobial activity against poultry pathogens. Yanang extract was prepared under different leaf processing conditions and used as a substrate for Pediococcus acidilactici V202, Lactiplantibacillus plantarum TISTR 926, Streptococcus thermophilus TISTR 894, Bacillus subtilis RP4-18, and Bacillus licheniformis 46-2. Fermentation at 37 °C for 24 h revealed that lactic acid bacteria (P. acidilactici V202, L. plantarum TISTR 926, S. thermophilus TISTR 894) reduced pH (<4.10, p < 0.001) while maintaining high viable counts (>8.67 log CFU/mL, p < 0.01), whereas Bacillus strains (B. subtilis RP4-18, B. licheniformis 46-2) retained a higher pH (>5.00) and lower viability (<8.50 log CFU/mL). Total soluble solids decreased across treatments, with the lowest observed for B. subtilis RP4-18 (1.97 °Brix, p = 0.007). Freeze-dried probiotics encapsulated in enzyme-extracted rice bran carriers had comparable physicochemical properties (p > 0.05), while compared with Bacillus strains (p < 0.01), lactic acid bacteria had superior tolerance to simulated gastrointestinal and thermal stress. Supernatant from Yanang extract inhibited B. cereus WU22001, S. aureus ATCC25923, Escherichia coli ATCC25922, and Salmonella typhimurium WU241001 (MIC/MBC 25–50% v/v). These results indicate that Yanang extract supports effective probiotic fermentation, and rice bran encapsulation enhances survivability and antimicrobial functionality for potential functional feed applications. Full article

Bacillus subtilis is applied as a biofertilizer, biocontrol agent, and probiotic in agriculture, and is also used for industrial synthesis of proteins and peptides. These applications can be combined by using B. subtilis to synthesize plant health-promoting peptides in culture or to deliver them to roots via seed treatments. To facilitate the use of B. subtilis as a cell factory, we tested different media, temperatures, and growth phases to optimize ectopic expression of a Plant Elicitor Peptide from soybean (GmPEP3) that enhances seedling growth. Our results indicate that temperature, culture media, and growth phase have interactive effects, and that 30 °C and 2x YT media can enhance ectopic expression per cell compared to 37 °C or LB media in log phase bacteria. We also identified tradeoffs between cell growth and ectopic expression levels per cell, with the log phase favoring high expression per cell but the stationary phase yielding higher cell numbers and consequently higher expression levels per unit of growth media. In addition, to facilitate B. subtilis seed treatments, we compared retention of spores versus vegetative cells with and without carboxymethylcellulose (CMC) to improve the viability of B. subtilis seed treatments. Our results indicated that retention of viable bacteria on B. subtilis-treated seeds could be increased by ~40% by using the adhesive polymer CMC, and shelf life could be extended from 24 h to at least 3 months by using endospores rather than vegetative cells. For B. subtilis expressing GmPEP3, endospores also had comparable plant-growth-promoting activity as vegetative cells. This establishes the bioactivity of spores and illustrates the potential benefits of using B. subtilis to deliver heterologous peptides. These results provide valuable insights for deploying B. subtilis for crop health. Full article

In recent years, as the number of cats has increased, the intestinal health of cats has receied increasing amounts of attention. Probiotics have positive effects on maintaining intestinal homeostasis. This study was conducted to investigate the effects of probiotics Bacillus licheniformis (B. licheniformis) and Bacillus subtilis (B. subtilis) on cat immunity, inflammation, antioxidants, intestinal barrier and microbiota, and serum metabolites. Thirty-six cats (over one year old, 3.48 ± 0.71 kg) were randomly divided into 3 groups and fed with a basal diet (CON group), a basal diet + B. licheniformis (BL group), and a basal diet + B. subtilis (BS Group). The experiment lasted 35 days. Fecal scoring indicates that B. licheniformis and B. subtilis can improve fecal scores. Serum analysis indicated that the addition of both substances increased levels of IgA, IgM, T-AOC, and SOD, while reducing levels of the pro-inflammatory factor TNF-α. Moreover, 16S rRNA gene sequencing revealed that B. licheniformis and B. subtilis altered the fecal microbiota composition, characterized by the elevated abundance of Bacillus. Adding B. licheniformis to the diet increased the level of Faecalibacterium and decreased the level of Mogibacterium. Serum metabolomics revealed that levels of L-Glycine and Sn-Glycero-3-phosphocholine exhibited marked elevation in both the BL and BS groups, respectively. Furthermore, the study demonstrated that differential metabolites in the BL group were mainly enriched in amino acid metabolism pathways, while those in the BS group were chiefly concentrated in lipid metabolism pathways. However, this study acknowledges the limitations of its exclusive use of Ragdoll cats and its 35-day intervention period. It highlights the need for future research involving diverse breeds and longer durations. Overall, the data highlight B. licheniformis and B. subtilis as cat nutritional supplements that improve immunity and maintain intestinal health. Full article

Micro- and nanoplastics (MPs and NPs) are recognized as emerging contaminants posing potential risks to human health. Recent evidence highlights the potential of food-grade microbial strains to bind these particles and facilitate their removal, suggesting a promising probiotic-based strategy for mitigating their adverse health effects. This study investigated the adsorption and biodegradation capabilities of Bacillus subtilis DCP04, a strain isolated from Korean fermented soybean paste, cheonggukjang, on low-density polyethylene (LDPE) particles. Biofilm formation assays and morphological observations confirmed the strain’s ability to adhere to the surface of LDPE. Subsequent experiments demonstrated that DCP04 effectively adsorbed LDPE particles in a size-, time-, and concentration-dependent manner. This interaction induced significant morphological changes and increased hydrophilicity on the polymer surface. Furthermore, a positive correlation was observed between the activities of laccase and manganese peroxidase and a measurable weight loss in LDPE films, suggesting direct enzymatic involvement in polymer degradation. Crucially, the DCP04 strain also met key safety and functional criteria for use as a probiotic. These findings highlight the potential of DCP04 strain as a functional probiotic agent for mitigating the accumulation of MPs and NPs within the human body. Full article

Olive flounder (Paralichthys olivaceus), a key aquaculture species in East Asia, is prone to stress and bacterial diseases under intensive farming. Antibiotics are often used to control these problems, but their overuse promotes resistance and threatens sustainability. To provide safer alternatives, this study evaluated the strain-specific effects of dietary probiotics on growth, immunity, and disease resistance in olive flounder. A five-week feeding trial was conducted to evaluate the effects of three isolates—Bacillus subtilis, Enterococcus faecium, and Pediococcus pentosaceus—on growth, blood biochemistry, immune responses, and resistance against Edwardsiella tarda. Each strain was incorporated individually into a basal diet. After the feeding trial, probiotic supplementation improved growth performance. P. pentosaceus significantly increased final body weight and other growth indices (p < 0.05), while E. faecium yielded the lowest feed conversion ratio. Plasma glucose was markedly reduced in the E. faecium and P. pentosaceus groups (p < 0.001), whereas other biochemical indices remained stable. Phagocytic activity was significantly increased in the B. subtilis (p < 0.05) and P. pentosaceus (p < 0.01) groups, while lysozyme activity was significantly elevated in the E. faecium (p < 0.01) and P. pentosaceus (p < 0.05) groups. Following the E. tarda challenge, survival improved in all probiotic-fed groups (22.5–28.9%) compared with the control (11.5%). These findings demonstrate complementary, strain-specific benefits: P. pentosaceus enhanced growth, E. faecium improved feed efficiency and disease resistance, and B. subtilis stimulated immune responses. Validation under farm conditions and exploration of multi-strain formulations are warranted to optimize probiotic use in olive flounder aquaculture. Full article

To investigate the impacts of dietary Bacillus-based probiotics and yeast-derived prebiotics on the hepatic transcriptome profile, 500 Hisex White laying hens were randomly allotted into five dietary treatments from 37 to 52 weeks of age: control; control + Bacillus subtilis; control + Bacillus subtilis and Bacillus licheniformis; control + Bacillus coagulans; and control + Saccharomyces cerevisiae yeast cell wall. Transcriptome analysis revealed a substantial number of differentially expressed genes exclusively between the control and prebiotic groups, identifying 2221 genes (FDR ≤ 0.05), with 980 genes upregulated (log₂ fold change 0.69 to 24.62) and 1241 downregulated (log₂ fold change −0.74 to −26.46). The top 10 upregulated KEGG pathways included protein export, glycerophospholipid metabolism, tryptophan metabolism, amino acid biosynthesis, alanine, aspartate, and glutamate metabolism, cofactor biosynthesis, propanoate metabolism, ABC transporters, 2-oxocarboxylic acid metabolism, and protein processing within the endoplasmic reticulum. In contrast, the most prominently downregulated pathways encompassed fructose and mannose metabolism, hedgehog signaling, PPAR signaling, Notch signaling, GnRH signaling, cell adhesion molecules, cytokine–cytokine receptor interactions, apelin signaling, glycosaminoglycan degradation, and RIG-I-like receptor signaling. These findings advance understanding of the hepatic transcriptomic response to yeast-derived prebiotics and identify key molecular pathways that could be targeted to enhance metabolic function in laying hens. Full article

Bacterial nanocellulose (BNC) has gained considerable interest over the last decade due to its unique properties and versatile applications. However, the low yield and the high production cost significantly limit its industrial scalability. The proposed study explores the isolation of new BNC producers from date palm sap and the use of date waste extract as a sustainable carbon source to improve BNC productivity. Results revealed three potential BNC producers identified as Komagataeibacter sp. IS20, Komagataeibacter sp. IS21, and Komagataeibacter sp. IS22 with production yield of 1.7 g/L, 0.8 g/L and 1.8 g/L, respectively, in Hestrin-Schramm (HS) medium. The biopolymer characterization indicated the presence of type I cellulose, a high thermal stability, and a highly dense network made of cellulose nanofibrils for all BNC samples. The isolate IS22, showing the highest productivity, was selected for an optimization procedure using a full factorial design with date waste extract as a carbon source. The BNC yield increased to 6.59 g/L using 4% date waste extract and 2% ethanol after 10 days of incubation compared to the standard media (1.8 g/L). Two probiotic strains, including Bacillus subtilis (BS), and Lactobacillus plantarum (LP) were successfully encapsulated into BNC matrix through a co-culture approach. The BNC-LP and BNC-BS composites showed antibacterial activity against Pseudomonas aeruginosa. BNC–probiotic composites have emerged as a promising strategy for the effective delivery of viable probiotics in a wide range of applications. Overall, this study supports the use of date waste extract as a sustainable carbon source to enhance BNC productivity and reduce the environmental footprint using a high-yielding producer (IS22). Furthermore, the produced BNC demonstrated promising potential as an efficient carrier matrix for probiotic delivery. Full article

This study evaluated the effects of dietary probiotic-fermented corn wet distillers grains (FCWDGs) on finishing pigs. Three Bacillus subtilis strains (CGMCC21218, CCTCC2022073, and CICC10275) were used to ferment corn wet distillers grains, yielding FCWDGs-1, FCWDGs-2, and FCWDGs-3. A total of 128 130-day-old Anqing six white pigs were randomly assigned to four groups: a control group and groups supplemented with 6% FCWDGs-1 (T1), FCWDGs-2 (T2), and FCWDGs-3 (T3). Over a 60-day trial, FCWDGs significantly enhanced growth performance, with T1 and T3 groups showing higher final weight and average daily gain (ADG) compared to the control (p < 0.05), and feed-to-gain ratios were reduced in all treatments (p < 0.05). Loin muscle depth (LMD) was significantly greater in all treatments (p < 0.05), and the lean meat percentage (LMP) was significantly higher in the T1 group (p < 0.05). Antioxidant activity (T-AOC, SOD, and GSH-Px) was enhanced in all treatments, with the highest values observed in the T1 group (p < 0.05). Notably, FCWDGs reduced heavy-metal residues (As, Pb, Cu) in muscle, liver, and kidney tissues, particularly in the T1 group. The results highlight the potential of Bacillus subtilis-fermented FCWDGs to enhance growth performance and carcass traits, and reduce heavy metal accumulation in pig tissues. Full article

This study aimed to assess whether dietary supplementation with probiotics could alleviate intestinal injury in lipopolysaccharide (LPS)-challenged piglets. Healthy weaned piglets were randomly allocated to four individual groups (n = 6): (1) a control group; (2) an LPS group; (3) an LPS + Lactobacillus group; and (4) an LPS + Bacillus group. The control and LPS groups received a basal diet, while the probiotic groups were provided with the same basal diet supplemented with 6 × 10⁶ cfu/g of Lactobacillus casei (L. casei) or a combination of Bacillus subtilis (B. subtilis) and Bacillus licheniformis (B. licheniformis) at a dosage of 3 × 10⁶ cfu/g, respectively. On day 31 of the trial, overnight-fasted piglets were killed following the administration of either LPS or 0.9% NaCl solution. Blood samples and intestinal tissues were obtained for further analysis several hours later. The results indicate that dietary supplementation with probiotics significantly exhibited health-promoting effects compared with the control group and effectively reduced LPS-induced histomorphological damage to the small intestine, impairments in barrier function, and dysregulated immune responses via modulation of enzyme activity and the expression of relevant genes, such as nuclear factor-kappa B (NF-κB), interleukin 4 (IL-4), interleukin 6 (IL-6), interleukin 10 (IL-10), claudin-1, nuclear-associatedantigenki-67 (Ki-67), and β-defensins-1 (pBD-1). Collectively, these results suggest that dietary supplementation with probiotics could alleviate LPS-induced intestinal injury by enhancing the immunity and anti-inflammatory responses in piglets. Our research provides a theoretical basis for the rational application of probiotics in the future. Full article

Constipation is a prevalent gastrointestinal disorder that significantly impairs quality of life. While pharmacological agents such as loperamide are widely used to induce constipation in experimental models, there is increasing interest in natural alternatives for alleviating intestinal dysfunction. In this study, we investigated the laxative effects of soybean powder fermented by Bacillus subtilis DKU_09 in a loperamide-induced rat model of constipation. The probiotic strain was isolated from cheonggukjang, a traditional Korean fermented soybean paste, and its identity was confirmed through 16S rRNA sequencing. Fermented soybean powder was characterized morphologically via scanning electron microscopy and chemically via HPLC to assess its isoflavone content. Rats were administered loperamide (5 mg/kg) for four days to induce constipation and were then treated with fermented soybean powder at doses of 100, 200, or 300 mg/kg. No pharmacological laxatives (e.g., PEG) were used as a positive control; instead, values from the treatment groups were compared with those from the loperamide-only constipation group. Key outcomes of fecal output, water content, colonic fecal retention, and gastrointestinal transit ratio were measured. The fermented product significantly improved stool frequency and moisture content, reduced colonic fecal retention, and restored gastrointestinal transit in a dose-dependent manner. Notably, the 300 mg/kg group demonstrated nearly complete recovery of fecal parameters without affecting body weight. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. These findings suggest that Bacillus subtilis-fermented soybean powder exerts synergistic laxative effects through the combined action of probiotic viability and fermentation-enhanced bioactive compounds such as aglycone isoflavones. This study supports the potential use of fermented soybean-based nutraceuticals as a natural and safe intervention for constipation and gastrointestinal dysregulation. Full article

Background/Objectives: Listeria monocytogenes is a foodborne pathogen of major public health concern due to its ability to invade host cells and cause severe illness. This study aimed to develop and validate a multi-tiered screening pipeline to identify Bacillus strains with probiotic potential against L. monocytogenes. Methods: A total of 26 Bacillus isolates were screened for antimicrobial activity, gastrointestinal resilience, and host cell adhesion. A fluorescence-based infection assay using mCherry-expressing HCT 116 cells was used to assess cytoprotection against L. monocytogenes NCTC 7973. Eight strains significantly improved host cell viability and were validated by quantification of intracellular CFU. Two top candidates were tested in a murine model of listeriosis. The genome of the lead strain was sequenced to evaluate safety and biosynthetic potential. Results: B. subtilis CECT 8266 completely inhibited intracellular replication of L. monocytogenes in HCT 116 cells, reducing bacterial recovery to undetectable levels. In vivo, it decreased splenic bacterial burden by approximately 6-fold. Genomic analysis revealed eight bacteriocin biosynthetic clusters and silent antibiotic resistance genes within predicted genomic islands, as determined by CARD and Alien Hunter analysis. The strain also demonstrated bile and acid tolerance, as well as strong adhesion to epithelial cells. Conclusions: The proposed pipeline enables efficient identification of probiotic Bacillus strains with intracellular protective activity. B. subtilis CECT 8266 is a promising candidate for translational applications in food safety or health due to its efficacy, resilience, and safety profile. Full article

Background/Objectives: The ongoing evolution of SARS-CoV-2 has highlighted the limitations of parenteral vaccines in preventing viral transmission, largely due to their failure to elicit robust mucosal immunity. Methods: Here, we evaluated an intranasal (IN) vaccine formulation consisting of recombinant receptor-binding domain (RBD) adsorbed onto human probiotic Bacillus subtilis DG101 spores. Results: In BALB/c mice, IN spore-RBD immunization induced strong systemic and mucosal humoral responses, including elevated specific IgG, IgM, and IgA levels in serum, bronchoalveolar lavage fluid (BALF), nasal-associated lymphoid tissue (NALT), and saliva. It further promoted mucosal B cell and T cell memory, along with a Th1/Tc1-skewed T cell response, characterized by increased IFN-γ-expressing CD4⁺ and CD8⁺ T cells in the lungs. Conclusions: All in all, these findings highlight the potential of intranasal vaccines adjuvanted with probiotic B. subtilis spores in inducing sterilizing immunity and limiting SARS-CoV-2 transmission. Full article

The objective of this study was to evaluate the effects of various feed additives on odor emissions, gut health, and stress responses in laying hens fed low-protein diets. Four commercially available functional feed additives (Bacillus subtilis, protease, saponin, and thyme-based essential oil) were selected for this study. A total of 288 Hy-Line brown laying hens aged 49 weeks were randomly fed on one of six experiment diets: a 16% standard crude protein diet, a 12% low-crude-protein (LCP) diet, and LCP diets supplemented with Bacillus-based probiotic, protease, saponin, or thyme-based essential oils prepared for 8 weeks. Each treatment had eight replicates with six birds per replicate. Lowering crude protein levels affected the laying performance, nitrogen balance, odor production (i.e., ammonia), and nutrient digestibility but did not alter eggshell quality or fecal short-chain fatty acids. Dietary additives added into the LCP diet did not affect the laying performance, egg qualities, and nitrogen balance but increased crude ash digestibility compared with the LCP-diet-fed laying hens. Branched-chain fatty acids tended to be higher in all laying hens fed low-CP diets, irrespective of feed additives. Notably, low vs. standard protein diets tended to increase yolk corticosterone levels, which is an indicator of stress responses in chickens. This low-CP-mediated increase in yolk corticosterone was partially decreased by 20.8–48.6% on average, depending on the additives used. Our study suggests that low-protein diets could effectively lower nitrogen excretion and odor emissions. However, adding dietary additives into low-protein diets has minimal effects on low-CP-diet-fed laying hens, which needs further studies to clarify the role of low-crude-protein diets and dietary additives in modulating hindgut fermentation via shaping the gut microbiota and stress responses of laying hens. Full article