Phytonutrient diet supplementation promotes beneficial Clostridia species and intestinal mucus secretion resulting in protection against enteric infection.

Plant extracts, or phytonutrients, are used in traditional medicine practices as supplements to enhance the immune system and gain resistance to various infectious diseases and are used in animal production as health promoting feed additives. To date, there are no studies that have assessed their mechanism of action and ability to alter mucosal immune responses in the intestine. We characterized the immunomodulatory function of six phytonutrients: anethol, carvacrol, cinnamaldehyde, eugenol, capsicum oleoresin and garlic extract. Mice were treated with each phytonutrient to assess changes to colonic gene expression and mucus production. All six phytonutrients showed variable changes in expression of innate immune genes in the colon. However only eugenol stimulated production of the inner mucus layer, a key mucosal barrier to microbes. The mechanism by which eugenol causes mucus layer thickening likely involves microbial stimulation as analysis of the intestinal microbiota composition showed eugenol treatment led to an increase in abundance of specific families within the Clostridiales order. Further, eugenol treatment confers colonization resistance to the enteric pathogen Citrobacter rodentium. These results suggest that eugenol acts to strengthen the mucosal barrier by increasing the thickness of the inner mucus layer, which protects against invading pathogens and disease.

A commensal symbiosis between Prevotella bivia and Peptostreptococcus anaerobius involves amino acids: potential significance to the pathogenesis of bacterial vaginosis.

In both batch and continuous culture, Peptostreptococcus anaerobius was able to grow in vaginal defined medium with Prevotella bivia, but not in pure culture. Growth of P. anaerobius was increased by 238% (P < 0.001) in peptone-supplemented vaginal defined medium conditioned by prior growth of P. bivia. Analysis of P. bivia culture supernatants showed a net accumulation of amino acids and subsequent growth of P. anaerobius in the conditioned supernatants resulted generally in amino acid utilization. Supplementation of peptone-supplemented vaginal defined medium with amino acids in concentrations similar to those available after prior growth with P. bivia were growth-stimulatory (246%, P=0.006) for P. anaerobius. Increased availability of amino acids by P. bivia is proposed as a mechanism to support the observed in vitro commensal symbiosis between P. bivia and P. anaerobius.

beta-Lactamase producing bacteria in adult periodontitis.

In 23 untreated adult periodontitis patients, the occurrence of beta-lactamase producing periodontal bacteria was determined. In addition to non-selective isolation media, selective isolation and growth of beta-lactamase positive subgingival bacterial species was carried out on blood agar plates supplemented with amoxicillin and plates with amoxicillin+clavulanic acid. Porphyromonas gingivalis, Prevotella intermedia, Actinobacillus actinomycetemcomitans, Peptostreptococcus micros, Fusobacterium nucleatum, Bacteroides forsythus and Campylobacter rectus isolates from the non-selective medium were tested for beta-lactamase activity by a nitrocefin disk method (DrySlide) and by a laboratory chromogenic nitrocefin-based test. Isolates from the amoxicillin plates that were absent on the amoxicillin/clavulanic acid plates were identified and tested for beta-lactamase production. Based on the non-selective plates, six of 23 P. intermedia isolates, 2 of 19 B. forsythus isolates and 3 of 23 F. nucleatum isolates were beta-lactamase positive. The beta-lactamase positive species Prevotella loescheii, Prevotella buccae, Prevotella buccalis and Actinomyces spp were recovered from the selective amoxicillin plates. beta-Lactamase positive subgingival species were recovered from 17 of 23 patients (74%) but usually comprised low proportions of the subgingival microbiota (range < 0.01-15%). Comparison of the DrySlide test and the nitrocefin-based laboratory test revealed full agreement of test results. beta-Lactamase activity in whole subgingival plaque was detected in 12 patient samples (52%). It was concluded that beta-lactamase activity in subgingival bacteria in adult periodontitis is a common feature. However, since the majority of the samples showed only low-level enzymatic activity, the clinical relevance of this observation with regard to therapy with unprotected enzyme-susceptible beta-lactams is uncertain, though failure on the other hand, is difficult to rule out when a mechanism of resistance is present. The majority of beta-lactamase positive strains was found among species of the Prevotella genus.

Peptostreptococcus productus strain that grows rapidly with CO as the energy source.

Anaerobic bacteria were enriched with a sewage digestor sludge inoculum and a mineral medium supplemented with B-vitamins and 0.05% yeast extract and with a 50% CO-30% N2-20% CO2 (2 atm [202 kPa]) gas phase. Microscopic observation revealed an abundance of gram-positive cocci, 1.0 by 1.4 micron, which occurred in pairs or chains. The coccus, strain U-1, was isolated by using roll tubes with CO as the energy source. Based on morphology, sugars fermented, fermentation products from glucose (H2, acetate, lactate, and succinate), and other features, strain U-1 was identified as Peptostreptococcus productus IIb (similar to the type strain). The doubling time with up to 50% CO was 1.5 h; acetate and CO2 were the major products. In addition, no significant change in the doubling time was observed with 90% CO. Some stock strains were also able to use CO, although not as well. Strain U-1 produced acetate during growth with H2-CO2. Other C1 compounds did not support growth. Most probable numbers of CO utilizers morphologically identical with strain U-1 were 7.5 X 10(6) and 1.1 X 10(5) cells per g for anaerobic digestor sludge and human feces, respectively.

Growth of Peptococcus and Peptostreptococcus: effect of variations of culture media on efficiency of recovery.

Reference strains and clinical isolates of Peptococcus and Peptostreptococcus spp. were evaluated for their growth response in supplemented thioglycolate-yeast extract media. Supplements used included various combinations of hemin, menadione, sodium bicarbonate, and Tween 80. Parallel studies were done to compare the efficiency of recovery of viable cells grown in thioglycolate-based media and Wilkins-Chalgren broth and agar. In addition, the effects of age of the medium and medium storage on viable cell yields for reference strains were determined. Reference strains grown in freshly prepared thioglycolate-yeast extract medium supplemented with sodium bicarbonate produced a 10-fold greater increase in the number of viable cells recovered after 24 h of incubation than did the same organisms cultivated in Wilkins-Chalgren medium. The efficiency of recovery of organisms when either mid-logarithmic- or mid-stationary-phase cells were used to prepare standardized inocula was similar. The results suggest that thioglycolate-yeast extract medium supplemented with sodium bicarbonate is more productive than Wilkins-Chalgren medium for the cultivation of anaerobic gram-positive cocci and may represent a suitable alternative for antimicrobial susceptibility testing of these organisms.