Ox Bile

BACKGROUND: The mechanisms leading to occlusion of plastic biliary stents (PBS) are not known. OBJECTIVE: To evaluate the impact of reducing duodenobiliary reflux on stent patency rate. DESIGN: A newly designed antireflux PBS (AR-PBS) was tested in vitro by using ox bile. A prospective randomized trial in human beings was conducted. SETTING: Tertiary medical center. PATIENTS: Patients with malignant bile-duct strictures were studied. INTERVENTIONS: A PBS or an AR-PBS stent was placed by using standard techniques, and the patients were followed at regular intervals. Patients presenting with stent occlusion underwent re-stent placement with either a PBS or a metal stent. MAIN OUTCOME MEASUREMENTS: In vitro: resistance to retrograde flow and comparison of the basal and peak antegrade flow pressures between the 2 stents. In vivo: stent patency rates, complications, and the efficacy of the stents in improving the liver test. RESULTS: The AR-PBS stent could withstand a retrograde pressure gradient of >320 mm Hg compared with <1 mm Hg for the PBS. Secondary to the siphon effect of the valve, the antegrade flow resistance offered by the AR-PBS was on the negative side for all flow rates compared with PBS (P < .001). The median patency of the AR-PBS in human studies was 145 days (range, 52-252 days) compared with 101 days (range, 41-210 days) for the PBS (P = .002). Both stents were equally effective in improving the liver test, and complication rates were similar in the 2 groups. LIMITATIONS: The occluded stents were not examined microscopically. CONCLUSIONS: The antireflux biliary stent remains patent for a longer time and hence duodenobiliary reflux may be contributing to stent occlusion.

The ban on antibiotic growth promoters has intensified the demand for precise alternatives to sustain growth and gut health in poultry. The present study aimed to evaluate the impact of isolated autophages against Enterococcus bacteria resistant to high concentrations of bile on lipid digestion and chicken performance. A total of 240 one-day-old Arian × Hy-Line W36 chicks were assigned to five equal groups replicated six times with eight chicks per cage in a completely randomized design. Data were subjected to one-way ANOVA, and differences among treatments were considered significant at P < 0.05. The treatments included a control group, phage group (receiving an oral phage dose of 108 PFU) per day, Enterococcus group (receiving an oral Enterococcus dose of 106 CFU), ox bile (0.4 g/kg), and virginiamycin antibiotics (0.2 g/kg). Results showed that phage against Enterococcus sp. improved the feed conversion ratio (FCR) like virginiamycin (2.87 and 2.84, respectively) with a significant difference from other treatments (P < 0.05). Ether extract digestibility (81.48% vs. 79.27%) and in vitro oil-emulsification index (13.0% vs. 4.2%) were significantly increased in the phage-treated group compared with the control (P < 0.05), indicating improved lipid utilization. Phage against Enterococcus sp. increased intestinal gram-negative bacteria counted on MacConkey agar compared with other groups (P < 0.05). In vitro oil-emulsification index perfectly mirrored the inhibition of bile salt hydrolase (BSH) activity in ileum digesta of antibiotics and bacteriophage-treated groups. It could be concluded that specific inhibition of Enterococcus bacteria with high bile hydrolase activity by bacteriophage can improve EE digestibility and performance like growth-stimulating antibiotics. Plain Language Summary: This study explored a natural way to improve chicken growth without using antibiotics. Researchers used bacteriophages viruses that attack specific bacteria to remove Enterococcus bacteria from the gut. These bacteria break down bile acids, which are important for digesting fats. By reducing Enterococcus, chickens digested fat more effectively and showed better growth, simil

Dairy products are traditional sources of potentially bactriocinogenic and probiotic microorganisms, especially lactic acid bacteria (LAB). This study aimed to isolate bacteriocin-producing LAB from Bulgarian feta cheese and assess their biopreservative, probiotic potential, and safety. Pediococcus acidilactici ST41VF was isolated and shown to inhibit multiple Listeria monocytogenes strains through the production of bacteriocin. This bacteriocin remained stable across a range of pH levels, temperatures, and chemical exposures relevant to pharmaceutical contexts. The highest bacteriocin concentration (6400 AU/mL) occurred during the stationary growth phase. Safety assessments revealed that ST41VF does not produce biogenic amines or gelatinase, lacks proteolytic and mucin-degrading activity, and exhibits γ-hemolytic behavior. It tolerated gastrointestinal simulations, thriving in various pH levels, salt concentrations, and ox-bile. The strain produced 4.998 g/L of d-lactic acid and 7.110 g/L of l-lactic acid, a relevant information regarding potential negative effects of d-lacid acid. Considering its robust antimicrobial activity, resilience, and safety profile, P. acidilactici ST41VF demonstrates strong potential as a biopreservative in food systems and a probiotic candidate for therapeutic applications. © 2025 Wiley‐VCH GmbH.

Biliary tract infections (BTIs) arise within a bile-rich environment that profoundly shapes microbial ecology and pathogen adaptation. Pseudomonas aeruginosa, a major opportunistic pathogen in nosocomial settings, exhibits remarkable physiological plasticity, that enable persistence in such challenging niches. However, the influence of bile on P. aeruginosa's adaptive responses and phage-host interactions remains largely unexplored. Here, we demonstrate that ox-bile imposes concentration-dependent stress on P. aeruginosa strain ZS-PA-35, indicative of host-derived selective pressure. Notably, ox-bile enhances biofilm formation and promotes swarming and twitching motilities while concurrently suppressing swimming motility. Moreover, ox-bile modulates phage susceptibility, likely through altered receptor expression: exposure to ox-bile sensitizes P. aeruginosa to the type IV pili (T4P)-dependent phage phipa2, whereas susceptibility to the lipopolysaccharide (LPS)-targeting phage phipa10 remains unchanged. Genome-wide mutagenesis identified resistance-conferring mutations affecting T4P structures, LPS biosynthesis, and associated regulatory pathways. Among these, phage-resistant mutants ΔpilT and ΔgalU retained high fitness under ox-bile stress, accompanied by enhanced swarming and swimming motilities. Furthermore, in a lysogenic context, ox-bile markedly suppressed prophage accumulation in the T4P-dependent strain ZS-PA-05. These findings reveal that bile acts as a critical environmental cue shaping both adaptive physiology and phage susceptibility in P. aeruginosa, with broad implications for microbiome dynamics and the development of phage-based therapies targeting bile-impacted infections. © 2025. The Author(s).

Cryptococcus neoformans (C. neoformans) is a capsulated yeast that enters the body through inhalation and migrates via the bloodstream to the central nervous system, causing cryptococcal meningitis. Diagnosis methods are culture, serology, and India ink staining, which require cerebrospinal fluid (CSF) or whole blood. Molecular methods are used for epidemiological studies and require expensive commercial DNA extraction kits. This study aimed to develop an economical in-house method for extracting C. neoformans DNA from whole blood. C. neoformans cells of varying McFarland standards were spiked into expired blood, then lysed using laboratory-prepared lysis buffer and ox-bile solution, followed by organic DNA extraction. Ordinary PCR targeting the CNAG 04922 gene was performed. To determine the limit of detection, serial dilutions of C. neoformans were made, and DNA extraction was performed on other parts cultured on yeast extract peptone dextrose agar to determine colony-forming units (CFU). The lysis buffer method successfully extracted DNA from as low as the average of 62 CFU in 0.9 mL of expired blood with superior quality and high yield compared to ox-bile. The lysis buffer method yielded higher DNA quality and quantity than ox-bile and detected low concentrations of C. neoformans in expired blood. This method presents a cost-effective alternative for molecular diagnosis in resource-limited settings.

This study aims to investigate the resistance of potential probiotic Bacillus species to various conditions in the gastrointestinal (GI) tract and their safety characteristics. MALDI-TOF MS identified all tested strains with a good safety score of ≥ 2.0; the strains demonstrated the capacity to pass through the Gl tract, exhibiting a reduction of > 6 log/CFU live cells. Furthermore, they exhibited varying survival rates in an acidic environment (pH 2.0-3.0) and the presence of Ox-Bile (1% w/v) (p < 0.05). Following exposure to pH 3.0 and Ox-Bile, the survival rate of Bacillus spp. ranged between 85.94% and 91.24% and 87.30% and 91.54%, respectively. The results of the in vitro experiments showed that the six Bacillus strains had comparable characteristics (e.g., tolerance to GI track enzyme, auto-aggregation ability) to the reference probiotic strain Lactiplantibacillus plantarum LA15. The auto-aggregation results of the B. clausii BA8 strain, which has demonstrated resistance to GI tract conditions, were also noteworthy. This strain showed 72.32% after 2 h and 74.55% at the end of 5 h. Most suitable for use as probiotic strains B. clausii BA8 and B. subtilis BA11, sequenced via Illumina NovaSeq, showed average nucleotide identity (ANI) values of 98.1% and 97.8%, respectively. The genome annotation of B. clausii and B. subtilis with Prokka revealed 4,498,248-4,215,606 bp genome length, 44%-43% GC content, and 110-26 contigs, respectively. B. clausii BA8 has been comprehensively characterized, is of low risk for human consumption, and has been recommended as a potential probiotic strain. However, further in vivo experimentation is required to confirm these findings. © 2025 The Author(s). Food Science & Nutrition published by Wiley Periodicals LLC.

BACKGROUND: The global probiotics dietary supplements market size is continuously growing. To overcome probiotics' health concerns, metabiotics are recognized as a safer alternative. Aiming to deal with the escalating antimicrobial resistance, the current work demonstrates synergistic metabiotic-antibiotic combinations against antibiotic-resistant pathogens. METHODS: The probiotic properties of lactic acid bacteria (LAB) strains isolated from 3 commercial dietary supplements were characterized in vitro. The combinations of the cell-free supernatants (CFS) of selected probiotic strains and conventional antibiotics against Staphylococcus aureus and Escherichia coli clinical isolates were evaluated using the time-kill assay. To our knowledge, the current literature lacks sufficient time-kill assay studies revealing the kinetics of such metabiotic-antibiotic combinations against S. aureus and E. coli. RESULTS: Four LAB strains isolated from dietary supplements as well as two reference strains were included in this study. The isolated LAB strains were identified by MALDI-TOF mass spectrometry as follows: P2: Lactobacillus acidophilus, P3: Lactiplantibacillus plantarum, P4: Lacticaseibacillus rhamnosus, and P5: Pediococcus acidilactici. The identification matched with that annotated by the manufacturers, except for P3. The tested strains could resist the acidic environment at pH 3. Excluding P2, the examined strains showed less than 1 log reduction in survivors upon the addition of reconstituted skimmed milk to pepsin at pH 2 and displayed an acceptable tolerance to 0.3% ox-bile. All the strains tolerated pancreatin. The hydrophobicity and autoaggregation capacities ranged between 7-92% and 36-66%, respectively. P2 was excluded owing to its inferior probiotic potential. Although the remaining strains showed excellent growth at 0.2% phenol, their growth was reduced at higher concentrations. L. plantarum and P. acidilactici strains possessed bile salt hydrolysis activity. The time-kill assay revealed promising synergistic activities of the combinations of CFS of L. rhamnosus P4 with either ceftazidime or gentamicin against E. coli and with only ceftazidime against S. au

Pediococcus pentosaceus 732, Lactococcus lactis subsp. lactis 431, and Lactococcus lactis 808, bacteriocinogenic strains previously isolated from kimchi and banana, were investigated for their safety, beneficial properties and in vitro inhibition of pathogens such as Listeria monocytogenes ATCC 15313 and Staphylococcus simulans KACC 13241 and Staphylococcus auricularis KACC 13252. The results of performed physiological, biochemical, and biomolecular tests suggest that these strains can be deemed safe, as no virulence genes were detected in their DNA. Notably, only the gad gene associated with GABA production was identified in the DNA isolated of Lc. lactis 808 and Lc. lactis subsp. lactis 431 strains. All tested LAB strains exhibited γ-hemolysins and were non-producers of gelatinase and biogenic amines, which suggested their safety potential. Additionally, they were relatively susceptible to antibiotics except for streptomycin, tobramycin, and vancomycin for Pd. pentosaceus 732. The growth of Pd. pentosaceus 732, Lc. lactis subsp. lactis 431, and Lc. lactis 808 and their survival were minimally affected by up to 3% ox bile and low pH (except pH 2.0 and 4.0). Moreover, these LAB strains were not inhibited by various commercial extracts as well as most of the tested medications tested in the study. They did not produce proteolytic enzymes but exhibited production of D/L-lactic acid and β-galactosidase. They were also hydrophilic. Furthermore, their survival in artificial saliva, gastric simulation, and enteric passage was measured followed by a challenge test to assess their ability to inhibit the selected oral pathogens in an oral saliva model conditions. © 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.

ETHNOPHARMACOLOGICAL RELEVANCE: Sanchen powder is a traditional Tibetan medicine comprising Bambusae Concretio Silicea, Carthami Flos, and Bovis Calculus Artifactus. Bambusae Concretio Silicea is the dried mass of secreted fluid in the stalks of Gramineae plants such as Bambusa textilis McClure or Schizostachyum chinense Rendle. Carthami Flos is the dried flower of Carthamus tinctorius L. in the Compositae plant. Bovis Calculus Artifactus is made from ox bile powder, cholic acid, hyodeoxycholic acid, taurine, bilirubin, cholesterol, and trace elements. Research has evidenced the antibacterial efficacy of Sanchen powder, albeit its active constituents for this effect are yet to be established. AIM OF THE STUDY: To investigate effective compounds, potential targets, and molecular mechanism of Sanchen powder for its antibacterial properties by using network pharmacology combined with in vitro validation, with the aims of observing the action of effective compounds in Sanchen powder and exploring new therapeutic strategies for antibacterial. MATERIALS AND METHODS: In this study, UPLC-Q-TOF-MS was utilized to identify the chemical composition in Sanchen powder and its blood-borne chemical ingredients post-oral intake. A network pharmacology analysis was used to establish the chemical compound in the blood following oral administration-target-disease network. The study aimed to identify antibacterial active ingredients, which were then subjected to molecular docking and pharmacodynamic experiments to verify their efficacy. RESULTS: The findings demonstrate that following oral administration, the blood contains seven key components of Sanchen powder, including bilirubin, glycochenodeoxycholic acid, glycocholic acid, taurocholic acid, phenylalanine, safflomin A, and tryptophan. Additionally, the network pharmacology and molecular docking study results indicate the potential antibacterial effects of bilirubin, glycocholic acid, and glycochenodeoxycholic acid. In vitro antibacterial experiments revealed that bilirubin, glycocholic acid, and glycochenodeoxycholic acid could restrict the growth of the Staphylococcus aureus cell membrane at a certain concentration. Moreover,

Bile represses Salmonella enterica serovar Typhimurium (S. Typhimurium) intestinal cell invasion, but it remains unclear which bile components and mechanisms are implicated. Previous studies reported that bile inhibits the RamR binding to the ramA promoter, resulting in ramA increased transcription, and that ramA overexpression is associated to decreased expression of type III secretion system 1 (TTSS-1) invasion genes and to impaired intestinal cell invasiveness in S. Typhimurium. In this study, we assessed the possible involvement of the ramRA multidrug efflux regulatory locus and individual bile salts in the bile-mediated repression of S. Typhimurium invasion, using Caco-2 intestinal epithelial cells and S. Typhimurium strain ATCC 14028s. Our results indicate that (i) major primary bile salts, chenodeoxycholate and its conjugated-derivative salts, cholate, and deoxycholate, activate ramA transcription in a RamR-dependent manner, and (ii) it results in repression of hilA, encoding the master activator of TTSS-1 genes, and as a consequence in the repression of cellular invasiveness. On the other hand, crude ox bile extract and cholate were also shown to repress the transcription of hilA independently of RamR, and to inhibit cell invasion independently of ramRA. Altogether, these data suggest that bile-mediated repression of S. Typhimurium invasion occurs through pleiotropic effects involving partly ramRA, as well as other unknown regulatory pathways. Bile components other than the bile salts used in this study might also participate in this phenomenon.

Probiotic microorganisms are increasing their interest today due to the benefits they provide to humans. Vinegar is the process of processing foods containing carbohydrates that can be fermented by acetic acid bacteria and yeasts. Hawthorn vinegar is also important in terms of amino acids, aromatic compounds, organic acids, vitamins and minerals it contains. Depending on the variety of microorganisms in it, the content of hawthorn vinegar changes, especially its biological activity. Bacteria were isolated from handmade hawthorn vinegar obtained in this study. After performing its genotypic characterization, it has been tested that it can grow in low pH environment, survive in artificial gastric and small intestinal fluid, survive against bile acids, surface adhesion characteristics, antibiotic susceptibility, adhesion, and degrade various cholesterol precursors. According to the results obtained, the studied isolate is Levilactobacillus brevis, it can reproduce best at pH 6.3, survives 72.22% in simulated gastric juice, 69.59% in small intestinal fluid, and 97% adhesion to HTC-116. Partially reproduces even in the presence of 2% ox-bile, surface hydrophobicity is 46.29% for n-hexadecane. It has been determined that it can degrade 4 different cholesterol precursors except for Sodium thioglycolate and is generally resistant to antibiotics except for CN30 and N30. Considering the experimental findings of Levilactobacillus brevis isolated from hawthorn vinegar for the first time, it can be said that Levilactobacillus brevis has probiotic properties. © 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.

Comment in Radiology. 2019 Aug;292(2):407-408. doi: 10.1148/radiol.2019191172. Background Previously reported dual-energy CT methods for detecting noncalcified gallstones have reduced accuracy for gallstones smaller than 9 mm. Purpose To develop a dual-energy CT method for differentiating isoattenuating gallstones from bile and compare it with previously reported dual-energy CT methods by using a prospective ex vivo phantom reader study. Materials and Methods From May 2017 to May 2018, gallstones were collected from 105 patients (34 men; mean age, 51 years; age range, 18-84 years) undergoing cholecystectomy and placed inside 120-mL vials containing ox bile. The vials were placed inside a water-filled phantom and were scanned with dual-layer dual-energy CT. Thirty isoattenuating gallstones (4.3-24.7 mm in diameter) were evaluated. Conventional CT images, virtual noncontrast images, and monoenergetic images at 200 and 40 keV were created. Segmented images were created by using a two-dimensional histogram of Compton and photoelectric attenuation. Six readers evaluated the presence of isoattenuating gallstones in each image. Intra- and interreader agreement was measured by using percentage agreement, diagnostic performance was evaluated by using mean area under the receiver operating characteristic curve (AUC) estimates and pairwise comparisons, and the agreement of gallstone sizes measured at pathologic examination with those measured on segmented images was compared by using Bland-Altman analysis. Results For all gallstones, segmented images provided the highest mean intrareader (88.1%) and interreader (88.2% and 93.6%) agreements for all readers and reading sessions and the highest overall AUC (0.99; 95% confidence interval [CI]: 0.97, 1.00; adjusted P < .02 for all). For gallstones larger than 9 mm, no significant difference was found between the segmented and monoenergetic AUCs (all P > .94, adjusted P > .05 for all). For gallstones measuring 9 mm or smaller, the segmented images had the highest overall AUC (0.99; 95% CI: 0.97, 1.00; adjusted P < .01 for all). The mean difference in stone sizes was -0.6 mm, with limits of agreement from 2.6 to -3.8 mm. Conclus

Patients with ileectomy can present with severe steatorrhea due to bile acid depletion. While exogenous bile acid would seem to be ideal therapy for this condition, it is not often used because it is thought that the bile acid would be malabsorbed and would enter the colon, producing a secretory diarrhea. This report describes a patient who had an ileectomy and partial right colon resection for Crohn's disease and then developed severe steatorrhea due to bile acid malabsorption. Although steatorrhea was reduced from 134 to 9 g/24 hr with chronic ingestion of ox bile extract, stool weight did not increase with ox bile (stool weight 669 g/24 hr before therapy and 507 g/24 hr after therapy). In this patient, oral bile acid supplementation reduced fat excretion markedly, but did not aggravate diarrhea, even though the colon was still present. This result may have been due to impaired solubility of bile acid in fecal fluid due to an acid pH or to binding of bile acid with particles of solid stool. Ingestion of ox bile extract does not necessarily increase stool weight in patients with defective fat absorption due to bile acid malabsorption and can be tried with caution in an effort to diminish steatorrhea.

INTRODUCTION: Rapid and accurate diagnosis of causative pathogens in mastitis would minimize the imprudent use of antibiotics and, therefore, reduce the spread of antimicrobial resistance. Whole genome sequencing offers a unique opportunity to study the microbial community and antimicrobial resistance (AMR) in mastitis. However, the complexity of milk samples and the presence of a high amount of host DNA in milk from infected udders often make this very challenging. METHODS: Here, we tested 24 bovine milk samples (18 mastitis and six non-mastitis) using four different commercial kits (Qiagens' DNeasy® PowerFood® Microbial, Norgens' Milk Bacterial DNA Isolation, and Molzyms' MolYsis™ Plus and Complete5) in combination with filtration, low-speed centrifugation, nuclease, and 10% bile extract of male bovine (Ox bile). Isolated DNA was quantified, checked for the presence/absence of host and pathogen using PCR and sequenced using MinION nanopore sequencing. Bioinformatics analysis was performed for taxonomic classification and antimicrobial resistance gene detection. RESULTS: The results showed that kits designed explicitly for bacterial DNA isolation from food and dairy matrices could not deplete/minimize host DNA. Following using MolYsis™ Complete 5 + 10% Ox bile + micrococcal nuclease combination, on average, 17% and 66.5% of reads were classified as bovine and Staphylococcus aureus reads, respectively. This combination also effectively enriched other mastitis pathogens, including Escherichia coli and Streptococcus dysgalactiae. Furthermore, using this approach, we identified important AMR genes such as Tet (A), Tet (38), fosB-Saur, and blaZ. We showed that even 40 min of the MinION run was enough for bacterial identification and detecting the first AMR gene. CONCLUSION: We implemented an effective method (sensitivity of 100% and specificity of 92.3%) for host DNA removal and bacterial DNA enrichment (both gram-negative and positive) directly from bovine mastitis milk. To the best of our knowledge, this is the first culture- and amplification-independent study using nanopore-based metagenomic sequencing for real-time detection of the pathogen (within 5 hours) and

Necrotic enteritis (NE), mainly induced by the pathogens of Clostridium perfringens and coccidia, causes huge economic losses with limited intervention options in the poultry industry. This study investigated the role of specific bile acids on NE development. Day-old broiler chicks were assigned to six groups: noninfected, NE, and NE with four bile diets of 0.32% chicken bile, 0.15% commercial ox bile, 0.15% lithocholic acid (LCA), or 0.15% deoxycholic acid (DCA). The birds were infected with Eimeria maxima at day 18 and C. perfringens at day 23 and 24. The infected birds developed clinical NE signs. The NE birds suffered severe ileitis with villus blunting, crypt hyperplasia, epithelial line disintegration, and massive immune cell infiltration, while DCA and LCA prevented the ileitis histopathology. NE induced severe body weight gain (BWG) loss, while only DCA prevented NE-induced BWG loss. Notably, DCA reduced the NE-induced inflammatory response and the colonization and invasion of C. perfringens compared to NE birds. Consistently, NE reduced the total bile acids in the ileal digesta, while dietary DCA and commercial bile restored it. Together, this study showed that DCA and LCA reduced NE histopathology, suggesting that secondary bile acids, but not total bile acid levels, play an essential role in controlling the enteritis.

PURPOSE: To evaluate the effects of stomach and duodenal fluid on enamel surfaces, simulating the action of refluxed liquid in patients with duodenogastric reflux. METHODS AND MATERIALS: Forty bovine incisors were used to obtain enamel fragments. Only half of the enamel surface was exposed to erosive challenges; the samples were then randomly divided into the following four groups (n = 10): G1: HCl; G2: HCl + pepsin; G3: HCl + ox bile + NaHCO3; and G4: HCl + pancreatin + NaHCO3. The specimens were placed in 37°C solutions, six times per day, for 20 s, over a period of 5 days and then analysed for morphology, surface roughness and the step formed on the dental enamel using confocal laser microscopy. The data were analysed using the Kruskal-Wallis and Dunn's test (p <0.05). RESULTS: Both analyses revealed a higher step and surface roughness for the G3 group (5.6 μm ± 1.69, 2.2 μm ± 1.61), which were statistically significant compared with the G1 and G2 groups (3.9 μm ± 1.5 μm; 1.0 μm ± 0.18; 3.7 μm ± 1.45; and 0.9 μm ± 0.12) (p <0.05); only the step in the G4 group (4.9 μm ± 1.8 μm) was similar to that of the G3 group (p >0.05). Morphological analysis showed greater structural loss in the G3 and G4 groups. CONCLUSIONS: Bile and pancreatin, in combination with hydrochloric acid, may promote a greater loss of structure, increased surface roughness and loss of enamel prismatic anatomy.

AIM: The purpose of this study was to know the genetic and biochemical identification of isolated lactic acid bacteria (LAB) from Balinese bovine (Bos sondaicus) intestinal waste, acidity, and ox bile salts and to inhibit the growth pathogen of Staphylococcus aureus and Escherichia coli and the potential of those isolated to improve nutrient value of wheat pollard as animal feed ingredient by fermentation process. MATERIALS AND METHODS: This research was divided into three stages. The first stage, isolated LAB were obtained from the bovine intestines at a slaughterhouse in Indonesia. Small intestinal samples were collected from 10 healthy Balinese beef cattle (B. sondaicus). The isolated LAB were identified by VITEK 2, polymerase chain reaction, and 16S rDNA. The basic local alignment search tool (BLAST) was performed to determine the phylogenetic tree. The second stage, the LAB were screened for their tolerance at pH 2, 3, and 4; bile salt, and antagonistic to enteric pathogen. In the third stage, to determine the potency of this isolate to increase nutrient content of wheat pollard by facultative anaerobe fermentation for 3 and 5 days. RESULTS: The result of the first stage showed that the isolate could be identified as Lactobacilluscasei WPL 315. The result of the second stage showed that the isolate tolerance to low pH (pH 2, pH 3, and pH4) for 90 min and 24 h, and this isolate had viability tolerance in 0.3% bile salt. The isolate can inhibit S. aureus and E. coli. The result of the third stage by proximate analysis showed that crude protein increased by 23.08% after fermentation, while crude fiber decreased by 61.24% on the level 0.5% L. casei subsp. WPL 315 in the 3-day fermentation. CONCLUSION: Based on the results, it showed that L. casei WPL 315 derived from indigenous intestinal Balinese beef cattle (B. sondaicus) has tolerant characteristic on acidity and ox bile salts, has antagonistic effect against E. coli and S. aureus, and has the ability to increase crude protein and decrease crude fiber content of wheat pollard. It would be interesting to determine whether the strain has a probiotic candidate.

Lactobacillus salivarius LI01, isolated from healthy humans, has demonstrated probiotic properties in the prevention and treatment of liver failure. Tolerance to bile stress is crucial to allow lactobacilli to survive in the gastrointestinal tract and exert their benefits. In this work, we used a Digital Gene Expression transcriptomic and iTRAQ LC-MS/MS proteomic approach to examine the characteristics of LI01 in response to bile stress. Using culture medium with or without 0.15% ox bile, 591 differentially transcribed genes and 347 differentially expressed proteins were detected in LI01. Overall, we found the bile resistance of LI01 to be based on a highly remodeled cell envelope and a reinforced bile efflux system rather than on the activity of bile salt hydrolases. Additionally, some differentially expressed genes related to regulatory systems, the general stress response and central metabolism processes, also play roles in stress sensing, bile-induced damage prevention and energy efficiency. Moreover, bile salts appear to enhance proteolysis and amino acid uptake (especially aromatic amino acids) by LI01, which may support the liver protection properties of this strain. Altogether, this study establishes a model of global response mechanism to bile stress in L. salivarius LI01. BIOLOGICAL SIGNIFICANCE: L. salivarius strain LI01 exhibits not only antibacterial and antifungal properties but also exerts a good health-promoting effect in acute liver failure. As a potential probiotic strain, the bile-tolerance trait of strain LI01 is important, though this has not yet been explored. In this study, an analysis based on DGE and iTRAQ was performed to investigate the gene expression in strain LI01 under bile stress at the mRNA and protein levels, respectively. To our knowledge, this work also represents the first combined transcriptomic and proteomic analysis of the bile stress response mechanism in L. salivarius.

BACKGROUND: Enteric fever remains an important cause of morbidity in many low-income countries and Salmonella Paratyphi A has emerged as the aetiological agent in an increasing proportion of cases. Lack of adequate diagnostics hinders early diagnosis and prompt treatment of both typhoid and paratyphoid but development of assays to identify paratyphoid has been particularly neglected. Here we describe the development of a rapid and sensitive blood culture PCR method for detection of Salmonella Paratyphi A from blood, potentially allowing for appropriate diagnosis and antimicrobial treatment to be initiated on the same day. METHODS: Venous blood samples from volunteers experimentally challenged orally with Salmonella Paratyphi A, who subsequently developed paratyphoid, were taken on the day of diagnosis; 10 ml for quantitative blood culture and automated blood culture, and 5 ml for blood culture PCR. In the latter assay, bacteria were grown in tryptone soy broth containing 2.4% ox bile and micrococcal nuclease for 5 hours (37°C) before bacterial DNA was isolated for PCR detection targeting the fliC-a gene of Salmonella Paratyphi A. RESULTS: An optimized broth containing 2.4% ox bile and micrococcal nuclease, as well as a PCR test was developed for a blood culture PCR assay of Salmonella Paratyphi A. The volunteers diagnosed with paratyphoid had a median bacterial burden of 1 (range 0.1-6.9) CFU/ml blood. All the blood culture PCR positive cases where a positive bacterial growth was shown by quantitative blood culture had a bacterial burden of ≥ 0.3 CFU/ ml blood. The blood culture PCR assay identified an equal number of positive cases as automated blood culture at higher bacterial loads (≥0.3 CFU/ml blood), but utilized only half the volume of specimens. CONCLUSIONS: The blood culture PCR method for detection of Salmonella Paratyphi A can be completed within 9 hours and offers the potential for same-day diagnosis of enteric fever. Using 5 ml blood, it exhibited a lower limit of detection equal to 0.3 CFU/ml blood, and it performed at least as well as automated blood culture at higher bacterial loads (≥0.3 CFU/ml blood) of clinical specimens despite using half the