Antimicrobial and anti-inflammatory activities of leaf extract of Valeriana wallichii DC.

Valeriana wallichii DC (Valerianaceae) is one of the most widely used traditional remedies for various complications associated with nervous system and digestion. No antimicrobial and anti-inflammatory studies have so far been carried out on the aerial parts of the plant. The present work was focused to evaluate the antimicrobial (antifungal and antibacterial) and anti-inflammatory properties of V. wallichii using reported methods. Chloroform fraction (VW-2) and hexane fraction (VW-3) exhibited significant activity against S. aureus and B. subtilus, respectively. The chloroform fraction (VW-2) showed significant activity against S. aureus with 0.27 mg/ml MIC, where 0.31 mg/ml MIC was deduced for VW-3 fraction against B. subtilus. VW-3 fraction was also found to be the most potent inhibitor of M. canis, showing 70% inhibition with an MIC value of 0.19 mg/ml. Considerable inhibitory activity was also observed for VW-2 and water fraction (VW-6) against M. canis and A. flavus. A remarkable anti-inflammatory like activity was observed for the crude extract at a dose of 200 mg/kg at all observed durations. Other doses of the sample also showed excellent activity. Looking to these results it may be concluded that V. wallichii may be a potential source for activity guided isolation of natural products with antimicrobial and anti-inflammatory-like properties.

Tellurite resistance and reduction by a Paenibacillus sp. isolated from heavy metal-contaminated sediment.

A gram-positive bacterium (designated as strain TeW) that is highly resistant to tellurite was isolated from sediment. The bacterium can grow in the presence of up to 2,000 micromol/L of potassium tellurite (K2TeO3). Reduction of K2TeO3 to tellurium was indicated by the blackening of the growth medium. No lag in growth was observed when cells unexposed to tellurite were transferred to the growth medium containing K2TeO3, indicating that resistance to tellurite was not inducible. Up to 50 and 90% of the metalloid oxyanion tellurite (TeO(3)(2-)) was removed from the medium by strain TeW during growth in nonstatic (shaking) and static (without shaking) conditions, respectively. The bacterium was identified as a Paenibacillus sp. according to its morphology, physiology, and 16S rDNA sequence homology.

Evaluation of culture media for Paenibacillus larvae applied to studies of antimicrobial activity.

This study was conducted to compare different liquid culture media for Paenibacillus larvae growth in order to find the best one to be used in studies on activity of antimicrobial substances, such as essential oils. P. larvae presented poor growth in usual broths such as Mueller-Hinton, commonly employed in antimicrobial activity assays. Growth in liquid media was evaluated using Paenibacillus larvae strains isolated from hives located in different geographical zones. The MYT medium (Mueller-Hinton broth, yeast extract and thiamine) was selected out of the eight liquid media analyzed, as it proved to be the most adequate due to its higher absorbance at 620 nm. The following mean values were obtained from the four P. larvae strains: 0.227 +/- 0.016 for the Cobo strain, 0.279 +/- 0.015 for La Plata strain, 0.758 +/- 0.020 for Mechongué strain and 0.244 +/- 0.0079 for Sierra de los Padres strain, respectively.

Evaluation of culture media for Paenibacillus larvae applied to studies of antimicrobial activity / Evaluación de medios de cultivo para el crecimiento de Paenibacillus larvae aplicables en estudios de actividad antimicrobiana

This study was conducted to compare different liquid culture media for Paenibacillus larvae growth in order to find the best one to be used in studies on activity of antimicrobial substances, such as essential oils. P. larvae presented poor growth in usual broths such as Mueller-Hinton, commonly employed in antimicrobial activity assays. Growth in liquid media was evaluated using Paenibacillus larvae strains isolated from hives located in different geographical zones. The MYT medium (Mueller-Hinton broth, yeast extract and thiamine) was selected out of the eight liquid media analyzed, as it proved to be the most adequate due to its higher absorbance at 620 nm. The following mean values were obtained from the four P. larvae strains: 0.227 ± 0.016 for the Cobo strain, 0.279 ± 0.015 for La Plata strain, 0.758 ± 0.020 for Mechongué strain and 0.244 ± 0.0079 for Sierra de los Padres strain, respectively.

Este trabajo está orientado a comparar diferentes medios de cultivo líquidos para el crecimiento de Paenibacillus larvae. El objetivo fue encontrar el más apropiado para utilizar en estudios de actividad antimicrobiana de diferentes sustancias, tales como aceites esenciales. P. larvae presenta un crecimiento débil en medios de cultivo como el Mueller-Hinton, comúnmente usado en ensayos de actividad antimicrobiana. Se evaluó el crecimiento en caldos de cultivo de cepas aisladas de colmenas ubicadas en diferentes zonas geográficas. De los ocho medios analizados, el MYT (Mueller-Hinton, extracto de levadura y tiamina) mostró ser el más apropiado, en éste se observó el mayor valor de absorbancia a 620 nm. Los valores obtenidos en promedio para los cuatro aislamientos de P. larvae evaluados fueron 0,227 ± 0,016 (cepa de Cobo); 0,279 ± 0,015 (cepa de La Plata); 0,758 ± 0,020 (cepa de Mechongué) y 0,244 ± 0,0079 (cepa de Sierra de los Padres).

Citrinin, a mycotoxin from Penicillium citrinum, plays a role in inducing motility of Paenibacillus polymyxa.

Paenibacillus polymyxa, a Gram-positive low-G+C spore-forming soil bacterium, belongs to the plant growth-promoting rhizobacteria. The swarming motility of P. polymyxa strain E681 was greatly induced by a secondary metabolite, citrinin, produced by Penicillium citrinum KCTC6549 in a dose-dependent manner at concentrations of 2.5-15.0 microg mL(-1) on tryptic soy agar plates containing 1.0% (w/v) agar. Flagellum staining showed that citrinin activated the production of flagella by P. polymyxa. This result was supported by reverse transcriptase-PCR analysis of gene expression, which showed increased transcriptional levels of sigD and hag homologues of P. polymyxa E681 in the presence of citrinin. The results presented here show that a mycotoxin, citrinin, has a newly identified function of inducing bacterial motility by transcriptional activation of related genes. This finding contributes to our understanding of the interactions between bacteria and fungal strains in nature.

Swarming and complex pattern formation in Paenibacillus vortex studied by imaging and tracking cells.

BACKGROUND: Swarming motility allows microorganisms to move rapidly over surfaces. The Gram-positive bacterium Paenibacillus vortex exhibits advanced cooperative motility on agar plates resulting in intricate colonial patterns with geometries that are highly sensitive to the environment. The cellular mechanisms that underpin the complex multicellular organization of such a simple organism are not well understood. RESULTS: Swarming by P. vortex was studied by real-time light microscopy, by in situ scanning electron microscopy and by tracking the spread of antibiotic-resistant cells within antibiotic-sensitive colonies. When swarming, P. vortex was found to be peritrichously flagellated. Swarming by the curved cells of P. vortex occurred on an extremely wide range of media and agar concentrations (0.3 to 2.2% w/v). At high agar concentrations (> 1% w/v) rotating colonies formed that could be detached from the main mass of cells by withdrawal of cells into the latter. On lower percentage agars, cells moved in an extended network composed of interconnected "snakes" with short-term collision avoidance and sensitivity to extracts from swarming cells. P. vortex formed single Petri dish-wide "supercolonies" with a colony-wide exchange of motile cells. Swarming cells were coupled by rapidly forming, reversible and non-rigid connections to form a loose raft, apparently connected via flagella. Inhibitors of swarming (p-Nitrophenylglycerol and Congo Red) were identified. Mitomycin C was used to trigger filamentation without inhibiting growth or swarming; this facilitated dissection of the detail of swarming. Mitomycin C treatment resulted in malcoordinated swarming and abortive side branch formation and a strong tendency by a subpopulation of the cells to form minimal rotating aggregates of only a few cells. CONCLUSION: P. vortex creates complex macroscopic colonies within which there is considerable reflux and movement and interaction of cells. Cell shape, flagellation, the aversion of cell masses to fuse and temporary connections between proximate cells to form rafts were all features of the swarming and rotation of cell aggregates. Vigorous vortex formation was social, i.e. required > 1 cell. This is the first detailed examination of the swarming behaviour of this bacterium at the cellular level.

Temperature, soluble solids and pH effect on Alicyclobacillus acidoterrestris viability in lemon juice concentrate.

Alicyclobacillus acidoterrestris is a thermoacidophilic, non-pathogenic, spore-forming bacterium detected in spoiled commercial pasteurized fruit juice. Apple, white grape and tomato are particularly susceptible. A. acidoterrestris spores are resistant to lemon juice pasteurization (2 min at 82 degrees C), and they can germinate and grow causing spoilage. This contamination is characterized by a medicinal or disinfectant smell attributed to guaiacol (o-dihydroxybenzene) production and other taint chemicals. The aim of this work was to study the influence of temperature (82, 86, 92 and 95 degrees C), total soluble solids (SS) (6.20, 9.8, 50 and 68 degrees Brix) and pH (2.28, 2.45, 2.80, 3.25, 3.5) on decimal reduction time (D) of the A. acidoterrestris in clarified and non-clarified concentrated lemon juice. Once D-value was determined, the resistance of A. acidoterrestris at the assayed temperatures was confirmed. SS and pH influence spore viability, because spore resistance increases with higher SS (50 degrees Brix 22 min 82 degrees C-68 degrees Brix 28 min 82 degrees C) and pH values (pH 2.28, 17 min-pH 4.00, 22 min). Bacterial growth was lower in clarified lemon juice, 26 min at 82 degrees C, than in non-clarified lemon juice, 51 min at 82 degrees C. Temperature was the parameter that had the greatest influence on the D value.

[Inhibition of Paenibacillus larvae employing a mixture of essential oils and thymol]. / Inhibición de Paenibacillus larvae empleando una mezcla de aceites esenciales y timol.

In vitro antimicrobial activity of a mixture of two essential oils and thymol against Paenibacillus larvae, causal agent of American Foulbrood (AFB), was evaluated. The essential oils were extracted from cinnamon (Cinnamomum zeylanicum) and thyme (Thymus vulgaris). The third component used, thymol, is the major component of the essential oil of thyme which contains 39.9% of thymol. Minimal inhibitory concentration (MIC) in Mueller-Hinton broth by the tube dilution method and minimal bactericide concentration (MBC) on MYPGP agar were evaluated. Thyme registered MIC values of 150-250 microg/ml and MBC values of 200-300 microg/ml, while the MIC and MBC values obtained for cinnamon were of 50-100 microg/ml and 100-125 microg/ml. Thymol showed similar MIC and MBC values of 100-150 microg/ml. No significant differences between the bacterial strains were detected, but significant differences between essential oils and thymol activity were registered (P<0,01). An inhibitory synergetic effect on AFB was observed reducing MIC and MBC values due to the use of a mixture of 62.5% of thyme, 12.5% of cinnamon and 25% of thymol.