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This study investigated the effects of a Lactobacillus paracasei KL1 and Lactobacillus plantarum subsp. plantarum Zhang-LL mixed probiotic on Salmonella-caused pullorosis in chicks. A total of 120 1-day-old Nongda no.3 dwarf chicks were randomly assigned to 4 treatments, with 6 replicates of 5 birds each. The treatments were blank group, Salmonella pullorum–infected group, probiotic treatment group, and probiotic prevention (PP) group. All birds (n = 90) except those in the blank group were infected with S. pullorum on day 4. On day 14, the BW, ADG, mortality, pathology of tissue, cecum colony count, immune organ indices, cecal mucosa secretory IgA, and cytokines were investigated. The results showed that the chicks infected with S. pullorum were depressed and their BW reduced. The PP group had the highest ADG and lowest mortality rate (0%), whereas the S. pullorum–infected group had 37.50% mortality rate and lowest ADG. Pathologic sections showed that the probiotic treatment group had minor lesions but the PP group had no lesions in the ileum, cecum, and liver. Cecal Lactobacillus counts was the highest (P 0.05) in the bursa of Fabricius, whereas in the PP group, all the immune organs were increased (P

The quorum-sensing regulation of class II bacteriocin (AcH) synthesis in Lactobacillus plantarum subsp. plantarum Zhang-LL was studied. No detectable inhibition zone was formed by the supernatant of L. plantarum subsp. plantarum Zhang-LL culture in skim milk (SM) with an inoculum size of 7 × 102 CFU/mL after incubation for 36 h. Hence, this culture system was used to investigate the induced regulation mechanism of bacteriocin production in L. plantarum subsp. plantarum Zhang-LL. Bacteriocin production by this bacterium in SM medium was induced by treatment with inactivated culture supernatant from de Man Rogosa Sharpe (MRS) medium (supernatant-MRS). Pediocin AcH encoded by the papA gene in a plasmid in strain Zhang-LL was the inducer present in supernatant-MRS. This is the first report of the role of pediocin AcH in the quorum-sensing regulation of class II bacteriocin synthesis. The mRNA of the papA, papB, papC, and papD genes involved in bacteriocin synthesis by strain Zhang-LL in SM medium was upregulated significantly after being induced by pediocin AcH. This study offers the first evidence that the ABT40_05745, ABT40_05750, and ABT40_11975 components of two-component systems in L. plantarum subsp. plantarum Zhang-LL are involved in the induced regulation of AcH bacteriocin production. HIGHLIGHTS

Plantaricin BM-1, a class IIa bacteriocin produced by Lactobacillus plantarum BM-1, shows obvious antibacterial activity against Escherichia coli. However, the mechanism underlying the action of class IIa bacteriocins against gram-negative bacteria remains to be explored. The purpose of this study was to investigate the role of YbfA, a DUF2517 domain-containing protein, in the response of Escherichia coli K12 to plantaricin BM-1. The growth curve experiment and MIC experiment showed that the sensitivity of E. coli to plantaricin BM-1 was decreased by a ybfA null mutation. Electron microscopy showed that the ybfA null mutation reduced the surface rupture and contraction caused by plantaricin BM-1, and mitigated the effect of plantaricin BM-1 on the morphology of the E. coli cell membrane. Proteomics analysis showed that 323 proteins were differentially expressed in E. coli lacking the ybfA gene (P < 0.05); 118 proteins were downregulated, and 205 proteins were upregulated. The metabolic pathways containing the upregulated proteins mainly included outer membrane proteins, integral components of the plasma membrane, regulation of cell motility, and regulation of locomotion. The metabolic pathways involving the downregulated proteins mainly included outer membrane protein glycine betaine transport, amino-acid betaine transport, and transmembrane signaling receptor activity. The results of the proteomics analysis showed that the protein expression of the BasS/BasR two-component system was significantly increased (P < 0.05). Moreover, the expression levels of downstream proteins regulated by this two-component system were also significantly increased, including DgkA, FliC, and MlaE, which are involved in cell membrane structure and function, and RT-qPCR also confirmed this result. The growth curve showed that the sensitivity of E. coli to plantaricin BM-1 was significantly increased due to deletion of the BasS/BasR two-component system. Thus, deletion of ybfA in E. coli can increase the expression of the BasS/BasR two-component system and positively regulate the structure and function of the cell membrane to reduce the sensitivity to plantaricin BM-1. This will help to explore the mechanism of action of class IIa bacteriocins against gram-negative bacteria.

In this study, an active antibacterial packaging film was developed by coating a polyethylene terephthalate/polyvinylidene chloride/retort casting polypropylene (PPR) plastic multilayer film with plantaricin BM‐1 and chitosan. The characteristics of the active packaging film and its antibacterial effect for chilled meat preservation were evaluated. Our results indicated that the barrier properties against oxygen were improved significantly and the tensile strength and the elongation at break were changed slightly. The active plantaricin film significantly (P

Bacillus subtilis SH21 was observed to produce an antifungal protein that inhibited the growth of F. solani. To purify this protein, ammonium sulfate precipitation, gel filtration chromatography, and ion-exchange chromatography were used. The purity of the purified product was 91.33% according to high-performance liquid chromatography results. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis and liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis revealed that the molecular weight of the protein is 30.72 kDa. The results of the LC–MS/MS analysis and a subsequent sequence-database search indicated that this protein was a chitosanase, and thus, we named it chitosanase SH21. Scanning and transmission electron microscopy revealed that chitosanase SH21 appeared to inhibit the growth of F. solani by causing hyphal ablation, distortion, or abnormalities, and cell-wall depression. The minimum inhibitory concentration of chitosanase SH21 against F. solani was 68 µg/mL. Subsequently, the corresponding gene was cloned and sequenced, and sequence analysis indicated an open reading frame of 831 bp. The predicted secondary structure indicated that chitosanase SH21 has a typical a-helix from the glycoside hydrolase (GH) 46 family. The tertiary structure shared 40% similarity with that of Streptomyces sp. N174. This study provides a theoretical basis for a topical cream against fungal infections in agriculture and a selection marker on fungi.

Controversial results were yielded in some studies on the effects of Lactobacillus on ulcerative colitis (UC). In this work, we evaluated the effect of live or heat-killed Lactobacillus plantarum Zhang-LL cells and cell supernatants on dextran sulfate sodium (DSS)-induced UC in rats. The tested subjects were gavaged to rats for 7 days, and then 5% was used DSS repeatedly to induce chronic UC. The results of a 42-day experiment, indicated consistently and significantly that Both living and heat-killed cells of L. plantarum Zhang-LL could down-regulate content of pro-inflammatory cytokines (TNF-α, INF-γ) in serum, reduce intestinal permeability by increasing the expression of tight junction proteins zo-1, claudin-1 and occludin in colon tissue, and maintain intestinal microbiota stability by increasing intestinal microbiota diversity to alleviate the symptoms of chronic UC in rats with 100% survival rate. Of concern is, the heat-killed cells of L. plantarum Zhang-LL were more effective against DSS-induced chronic UC in rats.

Plantaricin BM-1 is a class IIa bacteriocin with a strong bactericidal effect on gram-positive bacteria. Although plantaricin BM-1 also inhibits the growth of some gram-negative bacteria, including Escherichia coli, the mechanism is not clear. In this study, we used tandem mass tag-based quantitative proteomics analysis to examine the inhibitory mechanism of plantaricin BM-1 against E. coli K12, and evaluated the morphological effects by electron microscopy. The results demonstrated that plantaricin BM-1 inhibits the growth of E. coli K12 by bacteriostatic action, mainly acting on the surface of the cell wall, leading to its collapse. Proteomic analysis identified 976 differentially expressed proteins (>1.2-fold change, p < 0.05) under treatment with plantaricin BM-1, including 490 up-regulated proteins and 486 down-regulated proteins. These proteins were mainly involved in peptidoglycan synthesis and energy metabolism pathways, including amino acid, glyoxylate and dicarboxylate, ABC transporter, and quorum-sensing pathways. Specifically, plantaricin BM-1 treatment significantly improved peptidoglycan synthesis and enhanced the tricarboxylic acid cycle in E. coli K12, and altered the expression of cell membrane proteins. These results provide new insight into the inhibition mechanism of class IIa bacteriocins on gram-negative bacteria, which can lay the foundation for its broader use as an alternative to conventional antibiotics.

Salmonella pullorum is a highly pathogenic bacteria in poultry industry. However, antibiotics were restricted in many countries because of the increasing risk of antibiotic resistance, Therefore, an environmental friendly and effective alternative strives to be developed. This study investigated the benefit of a probiotic-fermented herbal blend on the growth performance and gut microbiota of newborn broilers infected with S. pullorum. A total of 120 one-day-old dwarf male chicks were randomly allotted to 4 treatment groups, each including 5 replicates of 6 chicks: negative control (NC), positive control (PC), herbal blend (HB), and probiotic-fermented herbal blend (PF). All birds (n = 90), except for those in the NC, were infected with S. pullorum (1.69 × 108 CFU) on day 1. On day 11, body weight (BW), mortality, tissue pathology, cecal colony counts, immune organ indices, cecal mucosa secretory immunoglobulin A (sIgA) concentrations, and cecal cytokine mRNA expression levels were investigated. No mortality was observed after the PF treatment, and less pathological condition was in the ileum, cecum, and liver of HB and PF. BW, average daily gain and average daily feed intake were significant higher in the HB group compared to the PC and were the highest in the PF (P < 0.05). HB treatment significantly increased cecal populations of Lactobacilli, and decreased cecal populations of Escherichia coli and Salmonella, but results were more pronounced in the PF group (P < 0.05). Both HB and PF treatments increased cecal mucosa sIgA compared with the PC (P < 0.05). Tumor necrosis factor alpha and interferon gamma were lowest (P < 0.05) and interleukin 4 was the highest (P < 0.05) in PF, which exhibited similar levels to the NC group. PF treatment significantly improved the development of the thymus and bursa in S. pullorum-infected chicks. In conclusion, PF treatment prevented death, improved growth performance, regulated intestinal flora and enhanced immune ability of in S. pullorum-infected with chicks.

Plantaricin BM-1, a typical IIa bacteriocin, is produced by Lactobacillus plantarum BM-1, which can be isolated from a traditionally fermented Chinese meat product. This bacteriocin exhibits perfect thermal stability and broad inhibitory activity against certain foodborne pathogens. In this study, we investigated the effect of plantaricin BM-1 combined with physicochemical treatments (sodium nitrite, heat treatment, ultrahigh pressure technology) on the control of Listeria monocytogenes populations in cooked ham. According to our results, the addition of sodium nitrite (0.075 or 0.15 mg/mL) significantly inhibited the growth of L. monocytogenes in broth, plantaricin BM-1 combined with heat treatment reduced the viable counts of L. monocytogenes more than plantaricin BM-1 alone, and plantaricin BM-1 combined with sodium nitrite and heat treatment had the greatest antibacterial effect in broth. However, the addition of sodium nitrite or heat treatment alone did not inhibit the growth of L. monocytogenes in cooked ham. The combined application of 5,120 arbitrary units per gram (AU/g) plantaricin BM-1 and 0.075 mg/g sodium nitrite reduced L. monocytogenes significantly more effectively (by 1.97 log CFU/g) than application of 5,120 AU/g plantaricin alone. Plantaricin BM-1 application combined with sodium nitrite and heat treatment led to viable counts of L. monocytogenes below the level of detection for 49 days of storage. Moreover, in cooked ham treated with 5,120 AU/g plantaricin BM-1, 0.075 mg/g sodium nitrite, and ultrahigh pressure technology (400 MPa for 5 min), viable counts of L. monocytogenes were reduced by 5.79 log CFU/g compared with control samples at the end of storage. In conclusion, the combination of plantaricin BM-1 with physicochemical treatments had a synergistic inhibitory effect on L. monocytogenes in cooked ham.

BACKGROUNDEarly detection of lung cancer to allow curative treatment remains challenging. Cell-free circulating tumor DNA (ctDNA) analysis may aid in malignancy assessment and early cancer diagnosis of lung nodules found in screening imagery.METHODSThe multi-center clinical study enrolled 192 patients with operable occupying lung diseases. Plasma ctDNA, white blood cell genomic DNA (gDNA) and tumor tissue gDNA of each patient were analyzed by ultra-deep sequencing to an average of 35,000X of the coding regions of 65 lung cancer-related genes.RESULTSThe cohort consists of a quarter of benign lung diseases and three quarters of cancer patients with all histopathology subtypes. 64% of the cancer patients is at Stage I. Gene mutations detection in tissue gDNA and plasma ctDNA results in a sensitivity of 91% and specificity of 88%. When ctDNA assay was used as the test, the sensitivity was 69% and specificity 96%. As for the lung cancer patients, the assay detected 63%, 83%, 94% and 100%, for Stage I, II, III and IV, respectively. In a linear discriminant analysis, combination of ctDNA, patient age and a panel of serum biomarkers boosted the overall sensitivity to 80% at a specificity of 99%. 29 out of the 65 genes harbored mutations in the lung cancer patients with the largest number found in TP53 (30% plasma and 62% tumor tissue samples) and EGFR (20% and 40%, respectively).CONCLUSIONPlasma ctDNA was analyzed in lung nodule assessment and early cancer detection while an algorithm combining clinical information enhanced the test performance.

Escherichia colican induce a group of stress-response proteins, including carbon starvation-induced lipoprotein (Slp), which is an outer membrane lipoprotein expressed in response to stressful environments. In this paper,slpnull mutantE. coliwere constructed by insertion of the group II intron, and then the growth sensitivity of theslpmutant strain was measured under 0.6% (vol/vol) hydrogen peroxide. The changes in resistance to hydrogen peroxide stress were investigated by detecting antioxidant activity and gene expression in theslpmutant strain. The results showed that deletion of theslpgene increased the sensitivity ofE. coliunder 0.6% (vol/vol) hydrogen peroxide oxidative stress. Analysis of the unique mapping rates from the transcriptome libraries revealed that four of thirteen remarkably up/down-regulated genes inE. coliwere involved in antioxidant enzymes after mutation of theslpgene. Mutation of theslpgene caused a significant increase in catalase activity, which contributed to an increase in glutathione peroxidase activity. ThekatGgene was activated by the OxyR regulator, which was activated directly by 0.6% (vol/vol) hydrogen peroxide, and HPI encoded bykatGwas induced against oxidative stress. Therefore, the carbon starvation-induced lipoprotein Slp regulates the expression of antioxidant enzymes and the transcriptional activator OxyR in response to the hydrogen peroxide environment, ensuring that cells are protected from hydrogen peroxide oxidative stress at the level of enzyme activity and gene expression.

To investigate the effects of Kluyveromyces marxianus M3 isolated from Tibetan mushrooms on diet-induced hypercholesterolemia in rats, female Wistar rats were fed a high-cholesterol diet (HCD) for 28 d to generate hyperlipidemic models. Hyperlipidemic rats were assigned to four groups, which were individually treated with three different dosages of K. marxianus M3+HCD or physiological saline+HCD via oral gavage for 28 d. The total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) levels in the serum and liver of the rats were measured using commercially available enzyme kits. In addition, the liver morphology was also examined using hematoxylin and eosin staining and optical microscopy. According to our results, the serum and liver TC, TG, LDL-C levels and atherogenic index (AI) were significantly decreased in rats orally administered K. marxianus M3 (p

Inducing at sublethal pHs can strengthen the survival rate of Bifidobacterium at the lethal pH. In this study, we determined the optimal inducing condition of Bifidobacterium and analyzed the effect of a low pH environment on fatty acid (FA) composition and thiol content in Bifidobacterium. The result indicates that inducing at pH 4.5 significantly improved the survival rate of Bifidobacterium at pH 3,5 and 3.0. After being induced at pH 4.5, the percentage of C18:1 increased significantly, while the percentage of C16:0 decreased significantly. The results suggest inducing treatment strengthened the membrane fluidity of Bifidobacterium. After being stressed at the lethal pH, the percentage of saturated FA increased significantly in both the control and induced cells, while the percentage of unsaturated FA decreased significantly. The results suggested that, to survive at the lethal pH, the cells strengthened the membrane rigidity and weakened the membrane fluidity to block the entrance of H+ by changing the FA composition in Bifidobacterium. In addition, after being stressed, the thiol content of the control cells was significantly lower than that of the induced cells. This study provided the first data proof that showed FA and thiol participated in response to low pH in Bifidobacterium.

The effects of the Kluyveromyces marxianus M3 strain, isolated from Tibetan mushrooms, on plasma lipids, egg cholesterol level, egg quality, and intestinal health of laying hens were evaluated. In total, 160 Beijing fatty laying hens (43 weeks old) were divided into four groups and fed a basal diet supplemented with 0%, 0.1%, 0.3%, or 0.5% freeze-dried K. marxianus M3 powder for four weeks. The results showed that yeast supplementation reduced serum total cholesterol (TC), triglyceride (TG), low density lipoprotein-cholesterol (LDL-C), and very low density lipoprotein-cholesterol (VLDL-C) levels (p

Two application methods were used to investigate the effect of plantaricin BM-1 on the control of Listeria monocytogenes and background spoilage bacteria in sliced vacuum-packaged cooked ham without the addition of any chemical preservatives, including sodium nitrite, during 35 days of storage at 4°C. Regardless of the application method, plantaricin BM-1 treatment (320, 640, or 1,280 arbitrary units [AU]/g of sliced cooked ham) significantly (P < 0.05) reduced the survival of L. monocytogenes (inoculated at 4 log CFU/g of sliced ham) compared with its survival in the control during the first 21 days of storage at 4°C. The inhibitory effect of plantaricin applied to the surface of the ham was significantly better than the same concentration of plantaricin incorporated into the cooked ham (P < 0.0001) during storage. Even 320 AU/g plantaricin applied to the surface exhibited greater inhibition of L. monocytogenes than 1,280 AU/g plantaricin incorporated into the cooked ham on days 1, 14, and 28. A level of 1,280 AU/g plantaricin applied to the surface of the ham reduced L. monocytogenes counts to below the detection limit from the 1st to the 21st day of storage at 4°C. Afterwards, L. monocytogenes was able to regrow, and the viable counts of L. monocytogenes at the end of storage reached 2.76 log CFU/g (6.11 log CFU/g lower than in the control). In the control ham, the counts of background spoilage bacteria increased gradually and surpassed the microbiological spoilage limitation level on the 21st day of storage. However, plantaricin BM-1 treatment significantly (P < 0.05) reduced the survival of background spoilage bacteria in ham compared with their survival in the control from day 21 to 35 of storage at 4°C. A level of 1,280 AU/g plantaricin incorporated into cooked ham was the most effective, reducing the count of background spoilage bacteria count from an initial 2.0 log CFU/g to 1.5 log CFU/g on day 7. This was then maintained for another 14 days and finally increased to 2.76 log CFU/g at the end of the storage at 4°C (2.85 log CFU/g lower than in the control). In conclusion, plantaricin BM-1 application inhibited the growth of L. monocytogenes and background spoilage bacteria in cooked ham during storage at 4°C and could be used as an antimicrobial additive for meat preservation.

The effect of plantaricin BM-1 on the behavior of Listeria Monocytogenes in sliced brined cooked ham during refrigerated storage (4°Cand 9°C) was assessed. The combination of low storage temperature (4°C) and plantaricin can significantly reduced viable counts of L. Monocytogenes during storage (P < 0.05). When the same amount of plantaricin was applied, the growth of L. Monocytogenes in brined cooked ham stored at 4 °C was significantly slower than that in sample stored at 9°C (P < 0.01). Even combination of 1280 AU/g plantaricin and 4°C storage produced greater inhibition of L. monocytogenes than combination of 5120 AU/g plantaricin and 9°C. The combination of 4°C and 5120 AU/g plantaricin was the most effective treatment and can decrease the levels of L. monocytogenes 1.7 Log CFU/g at the end of storage. Keywords-component; refrigerated storage; plantaricin; Listeria Monocytogenes; brined cooked ham

In recent years, bacteriocin as a natural antimicrobial compound, provides enormous promise to be used in food safety preservation. In this work, the polyethylene(PE)-based biological preservative films incorporating plantaricin BM-1, a typical IIa bacteriocin produced by Lactobacillus plantarum BM-1, were developed and characterized . The results showed that polyethylene (PE), low-density polyethylene (LDPE) and high-density polyethylene (HDPE) films soaked in plantaricin BM-1 solution had obvious antimicrobial activities aganist Listeria monocytogenes . And the volume of plantaricin BM-1 solution absorbed by PE, LDPE and HDPE films continued to increase and reached the maximum during exposure for up to 10, 6 and 16 hours, respectively. And the maximum absorption volumes of plantaricin BM-1 solution had no significant difference ( p >0.05) between the PE, LDPE and HDPE films. When soaking in water, the release amount of plantaricin BM-1 from active PE, LDPE and HDPE films reached the maximum potency at 16, 12 and 20 hours respectively. And the maximum release amount of plantaricin BM-1 from PE and LDPE active films were dramatically more than the HDPE active film ( p