In vivo model of Propionibacterium (Cutibacterium) spp. biofilm in Drosophila melanogaster.

OBJECTIVES: Acne vulgaris is a common inflammatory disorder of the pilosebaceous unit and Propionibacterium acnes biofilm-forming ability is believed to be a contributing factor to the disease development. In vivo models mimicking hair follicle environment are lacking. The aim of this study was to develop an in vivo Propionibacterium spp. biofilm model in Drosophila melanogaster (fruit fly). METHODS: We created a sterile line of D. melanogaster able to sustain Propionibacterium spp. biofilms in the gut. In order to mimic the lipid-rich, anaerobic environment of the hair follicle, fruit flies were maintained on lipid-rich diet. Propionibacterium spp. biofilms were visualized by immunofluorescence and scanning electron microscopy. We further tested if the biofilm-dispersal activity of DNase I can be demonstrated in the developed model. RESULTS: We have demonstrated the feasibility of our in vivo model for development and study of P. acnes, P. granulosum and P. avidum biofilms. The model is suitable to evaluate dispersal as well as other agents against P. acnes biofilm. CONCLUSIONS: We report a novel in vivo model for studying Propionibacterium spp. biofilms. The model can be suitable for both mechanistic as well as interventional studies.

Neonatal intestinal immune regulation by the commensal bacterium, P. UF1.

Newborns are highly susceptible to pathogenic infections with significant worldwide morbidity possibly due to an immature immune system. Recently, we reported that Propionibacterium strain, P. UF1, isolated from the gut microbiota of preterm infants, induced the differentiation of bacteria-specific Th17 cells. Here, we demonstrate that P. UF1 significantly increased the number of protective Th17 cells and maintained IL-10+ regulatory T cells (Tregs) in newborn mice. In addition, P. UF1 protected mice from intestinal Listeria monocytogenes (L. m) infection. P. UF1 also functionally sustained the gut microbiota and induced critical B vitamin metabolites implicated in the regulation of T cell immunity during L. m intestinal infection. Transcriptomic analysis of P. UF1-induced Th17 cells revealed genes involved in the differentiation and regulation of these cells. These results illustrate the potency of P. UF1 in the enhancement of neonatal host defense against intestinal pathogen infection.

A low-power ultrasound attenuation improves the stability of biofilm and hydrophobicity of Propionibacterium freudenreichii subsp. freudenreichii DSM 20271 and Acidipropionibacterium jensenii DSM 20535.

The main topic of this paper was to study the effect of ultrasound-attenuation (US) on the surface properties of propionibacteria (Acidipropionibacterium jensenii DSM 20535 and Propionibacterium freudenreichii DSM 20271). A preliminary screening was done by using different power levels (40 and 60%) and treatment times (4, 6, and 8 min); immediately after sonication, acidification and viable count were tested. The best combinations to avoid post-acidification after 6 h were the following: A. jensenii DSM 20535: power, 40%; time, 8 min; P. freudenreichii subsp. freudenreichii DSM 20271: power, 60%; time, 4 min. Moreover, the effect of US on the growth patterns, surface properties (biofilm formation and hydrophobicity), resistance to some selected antibiotics, and release of intracellular components was evaluated; the experiments were done immediately after the treatment. US-treatment improved the stability of biofilm after 5-7 days, caused an increase of hydrophobicity (from 15 to 27%) immediately after sonication, and determined an increase of cell permeability, as suggested by the release of intracellular components within 24 h and by the increased sensitivity to some antibiotics. This paper is the first report on US-attenuation on propionibacteria and could the background for future researches to modulate the surface properties of these microorganisms.

Clinical and Biological Features of Cutibacterium (Formerly Propionibacterium) avidum, an Underrecognized Microorganism.

The recent description of the genus Cutibacterium has altered the taxonomy of Propionibacterium species. These organisms still belong to the genera of the skin coryneform group, and the most-studied species remains Cutibacterium acnes. Cutibacterium avidum is also a known skin commensal. This underrecognized microorganism can, however, act as a pathogen after bacterial seeding and can be considered opportunistic, causing either superficial or deep/invasive infections. It can cause numerous infections, including but not limited to breast infections, skin abscesses, infective endocarditis, and device-related infections. The ecological niche of C. avidum is clearly different from that of other members of the genus: it is found in the axillary region or at wet sites rather than in dry, exposed areas, and the number of microorganisms increases during puberty. Historically, it has been used for its ability to modulate the immune response and for its antitumor properties. Conventional microbial culture methods and identification processes allow for its accurate identification and characterization. Thanks to the modern omics tools used for phylogenomic approaches, understanding C. avidum pathogenesis (including host-bacterium interactions and virulence factor characterization) is becoming easier, allowing for more thorough molecular characterization. These analyses have revealed that C. avidum causes diverse diseases mediated by multiple virulence factors. The recent genome approach has revealed specific genomic regions within this species that are involved in adherence and biofilm formation as well as fitness, survival, and defense functions. Numerous regions show the presence of phages and horizontal gene transfer. C. avidum remains highly sensitive to a broad spectrum of antibiotics, such as ß-lactams, fluoroquinolones, macrolides, and rifampin, although erythromycin and clindamycin resistance has been described. A long-term treatment regimen with a combination of antibiotics is required to successfully eliminate the remaining adherent bacteria, particularly in the case of deep infections after debridement surgery.

Comparative genomics and transcriptomics analysis-guided metabolic engineering of Propionibacterium acidipropionici for improved propionic acid production.

Acid stress induced by the accumulation of organic acids during the fermentation of propionibacteria is a severe limitation in the microbial production of propionic acid (PA). To enhance the acid resistance of strains, the tolerance mechanisms of cells must first be understood. In this study, comparative genomic and transcriptomic analyses were conducted on wild-type and acid-tolerant Propionibacterium acidipropionici to reveal the microbial response of cells to acid stress during fermentation. Combined with the results of previous proteomic and metabolomic studies, several potential acid-resistance mechanisms of P. acidipropionici were analyzed. Energy metabolism and transporter activity of cells were regulated to maintain pH homeostasis by balancing transmembrane transport of protons and ions; redundant protons were eliminated by enhancing the metabolism of certain amino acids for a relatively stable intracellular microenvironment; and protective mechanism of macromolecules were also induced to repair damage to proteins and DNA by acids. Transcriptomic data indicated that the synthesis of acetate and lactate were undesirable in the acid-resistant mutant, the expression of which was 2.21-fold downregulated. In addition, metabolomic data suggested that the accumulation of lactic acid and acetic acid reduced the carbon flow to PA and led to a decrease in pH. On this basis, we propose a metabolic engineering strategy to regulate the synthesis of lactic acid and acetic acid that will reduce by-products significantly and increase the PA yield by 12.2% to 10.31 ± 0.84 g/g DCW. Results of this study provide valuable guidance to understand the response of bacteria to acid stress and to construct microbial cell factories to produce organic acids by combining systems biology technologies with synthetic biology tools.

Revealing extracellular electron transfer mediated parasitism: energetic considerations.

Extracellular electron transfer (EET) is a mechanism that allows energetic coupling between two microorganisms or between a microorganism and an electrode surface. EET is either supported by direct physical contacts or mediated by electron shuttles. So far, studies dealing with interspecies EET (so-called IET) have mainly focused on possible syntrophic interactions between microorganisms favoured by this mechanism. In this article, the case of fermentative bacteria receiving extracellular electrons while fermenting a substrate is considered. A thermodynamical analysis based on metabolic energy balances was applied to re-investigate experimental data from the literature. Results suggest that the observations of a decrease of cell biomass yields of fermentative electron-accepting species, as mostly reported, can be unravelled by EET energetics and correspond to parasitism in case of IET. As an illustration, the growth yield decrease of Propionibacterium freudenreichii (-14%) observed in electro-fermentation experiments was fully explained by EET energetics when electrons were used by this species at a potential of -0.12 ± 0.01 V vs SHE. Analysis of other cases showed that, in addition to EET energetics in Clostridium pasteurianum, biological regulations can also be involved in such biomass yield decrease (-33% to -38%). Interestingly, the diminution of bacterial biomass production is always concomitant with an increased production of reduced compounds making IET-mediated parasitism and electro-fermentation attractive ways to optimize carbon fluxes in fermentation processes.

Potential influence of dairy propionibacteria on the growth and acid metabolism of Streptococcus bovis and Megasphaera elsdenii.

Ruminal acidosis is a prevalent disorder among dairy cows and feedlot cattle, which can significantly impair their health and productivity. This study, involving seven different strains of dairy propionibacteria, represents an in vitro investigation of the feasibility of using these organisms as direct-fed microbials to control lactic acid acumulation in the rumen. Interactions between the propionibacteria, Streptococcus bovis and Megasphaera elsdenii were evaluated in terms of effects on lactic, acetic and propionic acid metabolism, following co-incubation. Spot resistance tests showed slight but varying degrees of growth inhibition by S. bovis among the propionibacteria, while no inhibition was observed between M. elsdenii and the different strains of dairy propionibacteria. In the co-culture experiments comprising S. bovis in nutrient broth, significant differences in pH and the levels of production of lactic, acetic and propionic acid, were observed between treatments following inoculation with various propionibacteria and/or M. elsdenii. In general, lactic acid concentrations at the end of the incubation were significantly lower in the cultures containing propionibacteria compared with cultures comprising either S. bovis only or S. bovis + M. elsdenii, although efficacy of lactate metabolism varied between species and strains. Moreover,the accumulation of acetic and propionic acid in the combined cultures, but not in the solo S. bovis culture, indicated that these compounds were produced as a result of the metabolism of lactic acid by the propionibacteria and M. elsdenii.

Dairy propionibacteria as probiotics: recent evidences.

Nowdays there is evidence that dairy propionibacteria display probiotic properties, which as yet have been underestimated. The aim of this paper is to review the recent highlights of data representing the probiotic potential of dairy propionibacteria, studied both by general selection criteria (useful for all probiotic potentials), and by more specific and innovative approach. Dairy propionibacteria show a robust nature, that makes them able to overcome technological hurdles, allowing their future use in various fermented probiotic foods. In addition to the general selection criteria for probiotics in areas such as food safety, technological and digestive stress tolerance, many potential health benefits have been recently described for dairy propionibacteria, including, production of several active molecules and adhesion capability, that can mean a steady action in modulation of microbiota and of metabolic activity in the gut; their impact on intestinal inflammation, modulation of the immune system, potential modulation of risk factors for cancer development modulation of intestinal absorption.

Inhibition of enteropathogens adhesion to human enterocyte-like HT-29 cells by a dairy strain of Propionibacterium acidipropionici.

Adhesion to the host intestinal mucosa is considered relevant for orally delivered probiotics as it prolongs their persistence in the gut and their health promoting effects. Classical propionibacteria are microorganisms of interest due to their role as dairy starters as well as for their functions as probiotics. Propionibacterium acidipropionici Q4, is a dairy strain isolated from a Swiss-type cheese made in Argentina that displays probiotic potential. In the present work we assessed the ability of this strain to adhere to the human enterocyte-like HT-29 cell line and to counteract the adhesion of two common human enteropathogens, such as Escherichia coli C3 and Salmonella Enteritidis 90/390. The results were compared with those obtained with the well-known probiotic Lactobacillus rhamnosus GG. P. acidipropionici Q4 showed a high adhesion capacity, even higher than the reference strain L. rhamnosus GG (42.3±4.4% and 36.2±2.3%, respectively), whereas adhesion of enteropathogens was significantly lower (25.2±2.2% for E. coli and 21.0±3.4% for S. Enteritidis). Propionibacteria as well as lactobacilli were able to inhibit by exclusion and competition the adherence of E. coli C3 and S. Enteritidis 90/390 whereas only L. rhamnosus GG displaced S. Enteritidis from HT-29 intestinal cells. Inhibition of pathogens by propionibacteria was not exerted by antimicrobials or coaggregation but was mainly due to exclusion by cell surface components, such as proteins and carbohydrates. The relevance of cell surface proteins (CSP) for preventing pathogens infection was confirmed by their concentration dependent effect observed for both pathogens: 100 µg/ml of CSP inhibited E. coli attachment almost as untreated propionibacteria, whereas it partially inhibited the attachment of S. Enteritidis. Results suggest that P. acidipropionici Q4 could be considered for the development of propionibacteria containing functional foods helpful in counteracting enteropathogen infection.

Feeding wet distillers grains plus solubles with and without a direct-fed microbial to determine performance, carcass characteristics, and fecal shedding of O157:H7 in feedlot heifers.

The inclusion of wet distillers grains plus solubles (WDGS) in feedlot diets has become a common practice in many regions of the United States due to the expanded production of byproducts and fluctuating corn prices related to ethanol production and other factors. In addition, societal concerns over the continued use of antimicrobials in agriculture production combined with an enhanced interest in disease and pathogen prevention in the food supply have led to an increased interest in use of direct-fed microbials (DFM) in growing and finishing cattle. Direct-fed microbials have been shown to improve ADG and feed efficiency, alter ruminal fermentation, and decrease fecal shedding of potential harmful pathogens in feedlot cattle in some experiments. The objective of this experiment was to evaluate the effects of WDGS inclusion with or without a DFM containing (1 × 10 cfu ∙ heifer ∙ d) combined with (1 × 10 cfu ∙ heifer ∙ d) on the performance, carcass characteristics, and O157:H7 shedding in feedlot heifers. In early August, 288 crossbred heifers (initial BW = 295 ± 28 kg) were assigned to 1 of 4 treatments (12 pens per treatment; 6 heifers per pen) in a randomized complete block design with a 2 × 2 factorial arrangement of treatments. Body weights and fecal grab samples were obtained at approximately 28-d intervals throughout the experiment. Across the feeding period, heifers fed 30% WDGS tended ( = 0.09) to have greater ADG and had greater carcass-adjusted ADG ( = 0.05) compared with heifers fed dry-rolled corn (DRC). Dry matter intake was not affected ( = 0.65) by diet, although carcass-adjusted G:F tended ( = 0.10) to be improved for heifers fed WDGS. Heifers fed 30% WDGS tended ( ≤ 0.10) to have greater fat thickness at the 12th rib, lower marbling scores, and higher yield grades. The inclusion of . combined with . in the diet had no effect ( > 0.10) on performance or carcass merit in the present experiment. The incidence of O157:H7 throughout the experiment was low, with only 18 positive samples across all sampling periods. Neither WDGS inclusion nor the inclusion of . combined with . in the diet had any effect ( > 0.10) on O157:H7 shedding in this experiment. Feeding 30% WDGS to feedlot heifers improved animal performance compared to the DRC-based control diet.

Incorporation of Propionibacteria in Fermented Milks as a Probiotic.

Propionibacteria are mainly found in dairy products and fermented milks but are found in other foods as well. Dairy propionibacteria have recently shown to exert potential probiotic activities such as production of propionic acid, vitamins, bacteriocins, essential enzymes, and other vital metabolites. Furthermore, stimulating the immune system and lowering the blood cholesterol level are some of their favorable effects. They have a wide spectrum of antimicrobial activities, inhibiting the growth of gram-positive and some gram-negative bacteria, as well as some yeasts and molds. At industrial scale, they are used in cheese making, especially Swiss (hard) cheeses, as dominant starter cultures. There is a rising trend to use propionibacteria in fermented milks as probiotic. The current paper reviews the characteristics of propionibacteria related to their use in fermented milks either as starter culture or probiotic, methods for the enumeration of propionibacteria, and their functional (in vivo) efficiency.

Immunomodulation properties of multi-species fermented milks.

Dairy propionibacteria (PAB) are used as a ripening starter in combination with Lactic acid bacteria (LAB) for dairy products such as Swiss-type cheese. LAB and PAB have also been studied for their probiotic properties but little is still known about their individual and/or synergistic beneficial effects within dairy matrices. In the context of a rising incidence of Inflammatory Bowel Diseases, it has become crucial to evaluate the immunomodulatory potential of bacteria ingested in large numbers via dairy products. We therefore selected different strains and combinations of technological LAB and PAB. We determined their immunomodulatory potential by IL-10 and IL-12 induction, in human peripheral blood mononuclear cells, on either single or mixed cultures, grown on laboratory medium or directly in milk. Milk was fermented with selected anti-inflammatory strains of LAB or PAB/LAB mixed cultures and the resulting bacterial fractions were also evaluated for these properties, together with starter viability and optimum technological aspects. The most promising fermented milks were evaluated in the context of TNBS- or DSS-induced colitis in mice. The improvement in inflammatory parameters evidenced an alleviation of colitis symptoms as a result of fermented milk consumption. This effect was clearly strain-dependent and modulated by growth within a fermented dairy product. These findings offer new tools and perspectives for the development of immunomodulatory fermented dairy products for targeted populations.

Metabolic engineering of acid resistance elements to improve acid resistance and propionic acid production of Propionibacterium jensenii.

Propionic acid (PA) and its salts are widely used in the food, pharmaceutical, and chemical industries. Microbial production of PA by propionibacteria is a typical product-inhibited process, and acid resistance is crucial in the improvement of PA titers and productivity. We previously identified two key acid resistance elements-the arginine deaminase and glutamate decarboxylase systems-that protect propionibacteria against PA stress by maintaining intracellular pH homeostasis. In this study, we attempted to improve the acid resistance and PA production of Propionibacterium jensenii ATCC 4868 by engineering these elements. Specifically, five genes (arcA, arcC, gadB, gdh, and ybaS) encoding components of the arginine deaminase and glutamate decarboxylase systems were overexpressed in P. jensenii. The activities of the five enzymes in the engineered strains were 26.7-489.0% higher than those in wild-type P. jensenii. The growth rates of the engineered strains decreased, whereas specific PA production increased significantly compared with those of the wild-type strain. Among the overexpressed genes, gadB (encoding glutamate decarboxylase) increased PA resistance and yield most effectively; the PA resistance of P. jensenii-gadB was more than 10-fold higher than that of the wild-type strain, and the production titer, yield, and conversion ratio of PA reached 10.81 g/L, 5.92 g/g cells, and 0.56 g/g glycerol, representing increases of 22.0%, 23.8%, and 21.7%, respectively. We also investigated the effects of introducing these acid resistance elements on the transcript levels of related enzymes. The results showed that the expression of genes in the engineered pathways affected the expression of the other genes. Additionally, the intracellular pools of amino acids were altered as different genes were overexpressed, which may further contribute to the enhanced PA production. This study provides an effective strategy for improving PA production in propionibacteria; this strategy may be useful for the production of other organic acids. Biotechnol. Bioeng. 2016;113: 1294-1304. © 2015 Wiley Periodicals, Inc.

Absence of adverse events in healthy individuals using probiotics--analysis of six randomised studies by one study group.

Consumption of live bacteria as probiotic supplements is increasing. There is, however, a lack of information on the safety of ingested probiotics. The main objective of this study was to investigate the adverse events (AEs) of specific probiotics (Lactobacillus rhamnosus GG (LGG) alone or LGG in combination with L. rhamnosus Lc705, Propionibacterium freudenreichii JS, Bifidobacterium lactis BB12, or Bifidobacterium breve 99) studied in six of our study groups' clinical trials, by analysing individual participant data. A secondary objective was to study AEs associated with the consumed probiotic species and mixtures in three specific categories; 'gastrointestinal disorders', 'respiratory, thoracic and mediastinal disorders' and 'infections and infestations'. Six randomised, double-blind, placebo-controlled clinical studies by our study group were included in this AE analysis (study population n=1,909). All AE data were classified according to the National Cancer Institute's Common Terminology Criteria for Adverse Events (CTCAE) v4.0. From the 26 CTCAE System Organ Classes, we identified AEs in 20 classes among 1,909 subjects. Probiotic ingestion did not result in statistically significant differences in AEs in different groups, when compared to placebo. A subgroup analysis of gastrointestinal, respiratory, thoracic and mediastinal disorders, infections and infestations, found no differences between the intervention groups or for different probiotic combinations (risk ratio (RR) = 0.97, 95% confidence interval (CI): 0.93-1.02, P=0.30; RR=0.99, 95% CI: 0.97-1.01, P=0.35; RR=0.99, 95% CI: 0.93-1.06, P=0.62, respectively). As a conclusion, ingestion of probiotic supplementations containing LGG alone, or LGG in combination with L. rhamnosus Lc705, P. freudenreichii JS, B. breve 99, or B. lactis BB12 did not seem to cause AEs in young and elderly subjects in this analysis.

Electricity generation by Propionibacterium freudenreichii in a mediatorless microbial fuel cell.

OBJECTIVE: To test Propionibacterium freudenreichii as a novel biocatalyst in a glycerol-oxidizing microbial fuel cell (MFC). RESULTS: Two strains, P. freudenreichii ssp. shermanii and P. freudenreichii ssp. freudenreichii, were screened as anodic biocatalysts and shown to produce power from glycerol in an MFC. Voltage was generated with and without resazurin in the medium, showing that both strains are exoelectrogenic. Polarization data showed that an MFC with P. freudenreichii ssp. shermanii reached a maximum open circuit voltage of 485 mV and a maximum power density of 14.9 mW m(-2). Glycerol consumption was about 50 % lower in MFCs than in fermentations, indicating a metabolic shift in the MFC environment. CONCLUSION: P. freudenreichii ssp. shermanii and P. freudenreichii ssp. freudenreichii were shown for the first time to act as exoelectrogenic anodic biocatalysts in MFCs.

Emmental Cheese Environment Enhances Propionibacterium freudenreichii Stress Tolerance.

Dairy propionibacteria are actinomycetales found in various fermented food products. The main species, Propionibacterium freudenreichii, is generally recognized as safe and used both as probiotic and as cheese starter. Its probiotic efficacy tightly depends on its tolerance towards digestive stresses, which can be largely modulated by the ingested delivery vehicle. Indeed, tolerance of this bacterium is enhanced when it is consumed within a fermented dairy product, compared to a dried probiotic preparation. We investigated both stress tolerance and protein neosynthesis upon growth in i) chemically defined or ii) aqueous phase of Emmental cheeses. Although the same final population level was reached in both media, a slower growth and an enhanced survival of CIRM BIA 1 strain of P. freudenreichii subsp. shermanii was observed in Emmental juice, compared to chemically defined medium. This was accompanied by differences in substrates used and products released as well as overexpression of various early stress adaptation proteins in Emmental juice, compared to chemically defined medium, implied in protein folding, in aspartate catabolism, in biosynthesis of valine, leucine and isoleucine, in pyruvate metabolism in citrate cycle, in the propionate metabolism, as well as in oxidoreductases. All these changes led to a higher digestive stress tolerance after growth in Emmental juice. Mechanisms of stress adaptation were induced in this environment, in accordance with enhanced survival. This opens perspectives for the use of hard and semi-hard cheeses as delivery vehicle for probiotics with enhanced efficacy.

Screening of Propionibacterium spp. for potential probiotic properties.

The main topic of this paper is the evaluation of adhesion of propionibacteria to IPEC-J2 cells and the survival at pH 2.5 and with 0.3% bile salts added, bioactivity towards pathogens and antibiotic resistance of Propionibacterium freudenreichii subsp. shermanii, Propionibacterium jensenii, Propionibacterium acidipropionici and Propionibacterium thoenii. Adhesion to IPEC-J2 cell lines was ca. 25-35% and significantly increased with CaCl2. Moreover, propionibacteria showed a reduction of cell count of ca. 0.5% at pH 2.5 after 3 h, whereas cell count increased after 24 h with bile salts; finally, they significantly inhibited Escherichia coli O157:H7.

Real-time quantitative reverse transcription-polymerase chain reaction to detect propionibacterial ribosomal RNA in the lymph nodes of Chinese patients with sarcoidosis.

The aim of this study was to investigate the diagnostic value of using the copy number of propionibacterial rRNA as a biomarker for sarcoidosis. Ribosomal RNA of Propionibacterium acnes and P. granulosum was measured by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) using formalin-fixed and paraffin-embedded tissue of lymph node biopsy from 65 Chinese patients with sarcoidosis, 45 with tuberculosis and 50 controls with other diseases (23 with non-specific lymphadenitis and 27 with mediastinal lymph node metastasis from lung cancer). The receiver operating characteristic (ROC) curve was analysed to determine an optimal cut-off value for diagnosis, and the diagnostic accuracy of the cut-off value was evaluated in additional tissue samples [24 patients with sarcoidosis and 22 with tuberculosis (TB)]. P. acnes or P. granulosum rRNA was detected in 48 of the 65 sarcoidosis samples but only in four of the 45 TB samples and three of the 50 control samples. Analysis of the ROC curve revealed that an optimal cut-off value of the copy number of propionibacterial rRNA for diagnosis of sarcoidosis was 50·5 copies/ml with a sensitivity and specificity of 73·8 and 92·6%, respectively. Based on the cut-off value, 19 of the 24 additional sarcoidosis samples exhibited positive P. acnes or P. granulosum, whereas only one of the 22 additional TB samples was positive, resulting in a sensitivity and specificity of 79·2 and 95·5%, respectively. These findings suggest that propionibacteria might be associated with sarcoidosis granulomatous inflammation. Detection of propionibacterial rRNA by RT-PCR might possibly distinguish sarcoidosis from TB.

Reduced Burden of Salmonella enterica in Bovine Subiliac Lymph Nodes Associated with Administration of a Direct-fed Microbial.

Despite effective food safety interventions within abattoirs, Salmonella enterica remains a common contaminant of raw ground beef. Research has recently implicated peripheral lymph nodes (PLNs) as a potential route by which Salmonella contaminates ground beef. This study examined the efficacy of using Lactobacillus animalis (formerly designated Lactobacillus acidophilus; NP51) and Propionibacterium freudenreichii (NP24), at 10(9) cfu/head/day, as a direct-fed microbial (DFM) in feedlot cattle diets to control Salmonella within PLNs. Two studies were conducted in which cattle were randomly allocated into either control or DFM treatment groups. Diets of treated cattle were supplemented with 10(9) cfu/head/day of the DFM, while control groups received no DFM supplementation. During slaughter at abattoirs, one subiliac lymph node (SLN) per carcass was collected from 627 carcasses from one study and 99 carcasses from the second study. Lymph nodes were cultured to estimate the presence and concentration of Salmonella. In the first study, effects of DFM supplementation varied across slaughter days. On the first and second slaughter days, prevalence was reduced by 50% (P = 0.0072) and 31% (P = 0.0093), respectively. No significant difference was observed on slaughter day three (P = 0.1766). In the second study, Salmonella was 82% less likely (P = 0.008) to be recovered from SLNs of treatment cattle. While a greater relative risk reduction was observed in the latter study, absolute risk reductions were similar across studies. A significant reduction in the concentration of Salmonella in SLNs (P < 0.0001) on a cfu/g and cfu/node basis was also observed in cattle administered NP51 and NP24 in the first study; in the second study, too few quantifiable SLNs were observed to facilitate meaningful comparisons. The results indicate that NP51 and NP24 supplementation may aid in reducing the prevalence and concentration of Salmonella in SLNs and, therefore, serve as an effective control measure to reduce Salmonella in ground beef products.

High cell density propionic acid fermentation with an acid tolerant strain of Propionibacterium acidipropionici.

Propionic acid is an important chemical with wide applications and its production via fermentation is of great interest. However, economic production of bio-based propionic acid requires high product titer, yield, and productivity in the fermentation. A highly efficient and stable high cell density (HCD) fermentation process with cell recycle by centrifugation was developed for propionic acid production from glucose using an acid-tolerant strain of Propionibacterium acidipropionici, which had a higher specific growth rate, productivity, and acid tolerance compared to the wild type ATCC 4875. The sequential batch HCD fermentation at pH 6.5 produced propionic acid at a high titer of ∼40 g/L and productivity of 2.98 g/L h, with a yield of ∼0.44 g/g. The product yield increased to 0.53-0.62 g/g at a lower pH of 5.0-5.5, which, however, decreased the productivity to 1.28 g/L h. A higher final propionic acid titer of >55 g/L with a productivity of 2.23 g/L h was obtained in fed-batch HCD fermentation at pH 6.5. A 3-stage simulated fed-batch process in serum bottles produced 49.2 g/L propionic acid with a yield of 0.53 g/g and productivity of 0.66 g/L h. These productivities, yields and propionic acid titers were among the highest ever obtained in free-cell propionic acid fermentation.