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Antimicrobial effectiveness of pine needle extract on foodborne illness bacteria
Abstract
Fresh pine needles were collected and extracted with 95% methanol and the extract was concentrated to determine its antimicrobial activity. The methanol extract had a considerable inhibitory effect on the tested bacteria, such as Escherichia coli O157:H7, Listeria monocytogenes, Salmonella typhimurium, and Staphylococcus aureus. The methanol extract of pine needles was further fractionated to chloroform, ethylacetate, butanol, and water fractions. Among these four fractions, the butanol and water fractions, which showed a relatively strong inhibitory effect on all of the tested bacteria, were purified and the minimum inhibitory concentration (MIC) was determined for each microorganism. The MIC ranged between 25 mg/ml and 45 mg/ml depending on the microorganism. The purified active fractions were applied to sterilized milk as a model food system to define the antimicrobial effectiveness and it was found that the antimicrobial activities in the water fractions were stronger than those in the butanol fractions.
... [12] Ethyl acetate, butanol, and water fractions of the methanol extract of Pinus densiflora needles had strong antimicrobial activities to both Gram-negative and Gram-positive bacteria (Salmonella typhimurium, E. coli, L. monocytogenes, S. aureus) as essential oils had. [13] Therefore, low-and non-volatile pine metabolites of a different polarity have a still undisclosed antimicrobial potential. ...
... General 1 H-and 13 C-NMR spectra were registered on an AVANCE 300 spectrometer (Bruker BioSpin, Rheinstetten, Germany; 300. 13 ...
Lipophilic extractive metabolites from needles and defoliated twigs of Pinus armandii and P. kwangtungensis were studied by GC‐MS. Needles of P. armandii contained predominantly 15‐ O ‐functionalized labdane type acids (anticopalic acid), fatty acids, nonacosan‐10‐ol, sterols, nonacosan‐10‐ol and sterol saponifiable esters, and acylglycerols. While P. kwangtungensis needles comprised no anticopalic acid, but more trinorlabdane (14,15,16‐trinor‐8(17)‐labdene‐13,19‐dioic acid) and other labdane type acids, nonacosan‐10‐ol and its saponifiable esters. The major compounds in the P. armandii defoliated twig extract were abietane and isopimarane type acids, fatty acids, sterols, labdanoids ( cis‐ abienol), cembranoids (isocembrol and 4‐ epi ‐isocembrol), saponifiable sterol esters, and acylglycerols. The same extract of P. kwangtungensis contained larger quantities of fatty acids, caryophyllene oxide, serratanoids, sterols, saponifiable sterol esters, and acylglycerols; but lesser amounts of abietane and isopimarane type acids, cis‐ abienol, and lacked cembranoids. Both twig and needle extracts of P. armandii and P. kwangtungensis , as well as the extracts’ fractions, significantly inhibited the growth of Gram‐negative bacteria Serratia marcescens with MIC of 0.1 mg ml ‐1 . While in most cases they slightly stimulated the growth of Gram‐positive bacteria Bacillus subtilis at the same concentrations. Thus, lipophilic extractive compounds from needles and defoliated twigs of both pines are prospective for the development of antiseptics against Gram‐negative bacteria.
... Tea polyphenols, healthy and safe plant preparations [62,63], are broad-spectrum antibacterial agents that had inhibition and killing effects on Staphyloccocus aureus and other pathogenic intestinal bacteria [64][65][66]. Pine bark and heart wood had antibacterial effects on Staphylococcus aureus, Enterococcus faecium, and Bacillus subtilis [67], and pine needle extract had antimicrobial effectiveness against foodborne illness-causing bacteria [68]. Therefore, the manufacturer believed that VenaZn deodorant could effectively inhibit the growth of the microorganisms that produced harmful gases and absorb, catalyze, oxidize and decompose the harmful gases in livestock houses. ...
In order to mitigate the concentration of NH3 and greenhouse gases (GHGs: CO2, N2O, CH4) in livestock houses, two experiments, one determining the ideal manure removal frequency by cleaning the feces from a livestock house once, twice, three, and four times a day, and one in which microbial deodorant and VenaZn deodorant were sprayed, were conducted in a rabbit breeding house. The NH3, CO2, N2O, and CH4 concentrations were monitored continuously with an Innova 1512 photoacoustic gas monitor during the experiments. The results were as follows: the manure removal frequency had a significant impact on the average concentrations of NH3, CO2, and CH4 in the rabbit house. Cleaning the feces in the rabbit breeding house two to three times a day significantly reduced the NH3 concentration, and, on the contrary, cleaning the feces four times a day increased the NH3 concentration in rabbit house; increasing the manure removal frequency significantly reduced the concentrations of CO2 and CH4 in the rabbit house. Considering the average concentrations of NH3, CO2, N2O, and CH4 in the rabbit house and economic cost, it was better to remove feces twice a day. The average NH3 and CO2 concentration declined significantly within 3 days in the summer and winter; the N2O concentration declined within 3 days in the summer but did not decline in the winter; and there was no effect on the CH4 concentration in the summer and in the winter after spraying the rabbit house with microbial deodorant. Therefore, it was better to spray microbial deodorant twice a week on Monday and Thursday to reduce the NH3, CO2, and N2O concentrations in rabbit houses. The NH3, CO2, N2O, and CH4 concentrations first showed a decreasing trend and then an increasing trend over 5 days in the summer and 7 days in the winter after VenaZn deodorant was sprayed in the rabbit house, and the NH3, CO2, N2O, and CH4 concentrations on day 3 and day 4 were significantly lower than they were on the other days.
A membrane-based oligonucleotide array was used to detect predominant bacterial species in human fecal samples. Digoxygenin-labeled 16S rDNA probes were generated by PCR from DNA that had been extracted from fecal samples or slurries. These probes were hybridized to an array of 120 oligonucleotides with sequences specific for 40 different bacterial species commonly found in human feces, followed by color development using an alkaline phosphatase-conjugated antibody and NBT/BCIP. Twenty of the species were detected by this method, but E. coli, which was present at ∼1 × 10 5 CFU per gram feces, was not detected. To improve the sensitivity of this assay, reverse transcriptase-PCR was used to generate probes from RNA extracted from fecal cultures. Coupled with a chemiluminescence detection method, this approach lowered the detection limit for E. coli from ∼1 × 10 6 to ≤1 × 10 5. These results indicate that the membrane-array method with reverse transcriptase-PCR and chemiluminescence detection can simultaneously identify bacterial species present in fecal samples at cell concentrations as low as ≤1 × 10 5 CFU per gram.
The growth-inhibiting effects of Pinus densiflora leaf-derived materials on nine human intestinal bacteria were investigated using the impregnated paper disk method, and their activities were compared with those of 13 commercially available terpenes. The biologically active constituent of the extract of P. densiflora leaf was characterized as the monoterpene (1R)-(+)-α-pinene by various spectroscopic analyses. Responses varied according to bacterial strain, chemicals, and dose. At 10 mg/disk, limonene and (1R)-(+)-α-pinene strongly inhibited the growth of Clostridium perfringens, Staphylococcus aureus, and Escherichia coli, without adverse effects on the growth of five lactic acid-bacteria (Bifidobacterium adolescentis, B. bifidum, B. longum, Lactobacillus acidophilus, and L. casei). Little or no inhibition against seven bacteria was observed with anethole, borneol, camphor, caryophyllene, 1,8-cineole, estragole, linalool, and α-terpineol. Structure-activity relationship revealed that (1R)-(+)-α-pinene had more growth-inhibiting activity against C. perfringens than (1R)-(+)-β-, (1S)-(-)-α-, and (1S)-(-)-β-pinenes. Furthermore, the growth-inhibition against L. casei was much more pronounced in (1R)-(+)-β- and (1S)-(-)-β-pinenes than (1R)-(+)-α- and (1S)-(-)-α-pinenes. These results indicate that the (+)-α form seems to be required against C. perfringens and β form against L. casei for growth-inhibiting activity. Morphologically, most strains of C. perfringens were damaged and disappeared at 5 and 2 mg/disk of (1R)-(+)-α-pinene. Morphological study revealed that (1R)-(+)-α-pinene had more growth-inhibiting activity against C. perfringens than (1R)-(+)-β-, (1S)-(-)-α-, and (1S)-(-)-β-pinenes. As naturally occurring growth-inhibiting agents, the Pinus leaf-derived materials described above could be useful preventive agents against diseases caused by harmful intestinal bacteria such as clostridia.
In this study, to confirm the physiological activity of the grasshopper (Oxya chinensis sinuosa) antioxidant activity, antimicrobial activity, thermal stability of the antimicrobial substance, pH stability, total polyphenol content of the adult grasshopper was measured. Antibacterial activity in accordance with the extraction solvent showed a strong antibacterial activity in a mixed solvent of ethanol for the E. coli strain. Antimicrobial activity in 40^{\circ}C hot air drying and a freezedried condition was the highest and there was no difference in the gender. Antibacterial substance was stable to heat and pH. Antioxidant activity of the grasshopper exhibited a high activity in the 50% and 70% ethanol extract. Total polyphenol content was the 12 ~17 mg / 100 g and there was not great difference according to the drying conditions and gender.
Aqueous extracts of garlic (Allium sativum) preparation were prepared after the samples were exposed to various heat treatments. A quantitative assessment of antimicrobial activities was carried out by determining the minimum inhibitory and microbicidal concentrations (MICs and MMCs) of the various extracts against some selected bacteria and fungi. The antimicrobial activity of garlic decreased as the heating temperature increased. This fact implies that alliinase may be the most critical rate-determinant to produce the activity when garlic is heated.
A fluorescent glucose analogue, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose (2-NBDG), which had previously been developed for the analysis of glucose uptake in living cells, was investigated to determine its biological activity on microorganisms. 2-NBDG did not show any inhibitory effect on growth of yeast cells and bacteria. In contrast, 2-NBDG exhibited strong inhibitory effects on filamentous fungal growth. The growth of filamentous fungi was completely inhibited, when 2-NBDG was supplemented as sole carbon source. The inhibitory effect was decreased by the addition of glucose in the test medium. Furthermore, 2-NBDG inhibited chitinase activity of Trichoderma sp. These results suggested that the inhibitory effects of 2-NBDG on filamentous fungi might be partially due to the inhibition of chitinase.
Hot water extracts of 141 indigenous plants were screened for antibacterial activity against 7 Gram (+) and 12 Gram (-) bacteria by agar diffusion method. Of the 141 plants tested, 55 showed antibacterial activity against one or more species of Gram (+) bacteria and especially extracts of Oenothera stricta Ledebour (Onagraceae), Quercus variabilis Blume (Fagaceae), Alnus firma (Betulaceae), Geranium thunbergii (Geraniaceae), Lagerstroemia indica (Lythraceae) showed broad spectrum antibacterial activity on over 6 strains. Of the 141 plants tested, 45 showed antibacterial activity against one or more species of Gram (-) bacteria and especially extracts of Oenothera stricta Ledebour (Onagraceae), Punica granatum (Lythraceae), Quercus variabilis Blume (Fagaceae), Schizandra chinensis Baillon (Magnoliaceae), Alnus firma (Betulaceae), Alnus hirsuta (Betulaceae), Magnolia kobus (Magnoliaceae) showed broad spectrum antibacterial activity on over 8 strains. The most active antibacterial plants were Oenothera stricta Ledebour (Onagraceae), Punica granatum (Lythraceae), Quercus variabilis Blume (Fagaceae), Schizandra chinensis Baillon (Magnoliaceae) which are edible as permitted by Korea Food & Drug Adminstration. With in-depth research, the hot water extracts of these plants can be applied as food preservatives and alternatives of antimicrobials for livestock feeds.
To define diversity of domestic radish, we analysis genetic relationship of anti-fungal protein genes from several domestic radish (Raphanus sativus L.) seeds. We have isolated from domestic radish (Baekwoon) anti-fungal protein named RAP[12]. In this report, we isolate RNAs and raw protein from radish seeds then, RT-PCR analysis was done with another known anti-fungal sequences of radish from Gene Bank/EMBL and anti-fun- gal, anti-yeast activity were done against Bot교tis cenerea, Saccharomyces cerevisiaeι Candida albicans with it's raw proteins. The anti-fungal activity was shown used all seeds but anti-yeast activity was shown only two seeds (Myungsan, Baekwoon). RT-PCR products (about 0.2 Kb) were not shown only two seeds. To identify the sequencing relationship of the domestic radish, we have cloned and sequenced RAP genes of the radish and analysis the sequence relationship with clustalw program. Thus we report the result that there are some different relationship between domestic radish and known other radish's anti- fungal protein[15].
A novel antibacteria metabolite, ARK42, was isolated from a xerophilic fungal strain K42, and identified as Aspergillus repens based on its morphological characteristics. The metabolite exhibited antibacterial activities towards Staphylococcus aureus, Bacillus cereus, and Pseudomonas aeruginosa, with MICs of 25, 12.5, and 3.125 μg/ml, respectively, and killed Pseudomonas aeruginosa with minimal bactericidal concentration (MBC) of 12.5 μg/ml. Furthermore, anticancer activities were demonstrated against human colon cancer DLD-1 and lung cancer LXFL529 cells with an IC50 of 10 and 1 μg/ml, respectively.
The leukemia inhibitory factor (LIF), which is a very expensive reagent, can be used to efficiently control the differentiation
of human embryonic stem (ES) cells at concentrations >1000units/ml for 6–7days. However, in supplement <500units/ml, most
ES cells differentiate within 3–4days in in vitro cultures. α-Pinene from Pinus densiflora S. and a polysaccharide (MW 25kDa) from A. gigas Nakai showed promising results as a substitute for LIF in cultivating ES cells. By adding both 0.5 (μg/ml) of α-pinene and
the polysaccharide, most of the ES cells could be maintained under undifferentiated conditions after adding only 100units/ml
of LIF. It was found that α-pinene can play a role in preventing the ES cells from differentiating and the polysaccharide
can be used to grow the ES cells. The results suggest that human ES cells can be maintained under undifferentiated conditions
by supplementing both plant extracts, which can result in a reduction in the amount of LIF needed.
The antibacterial effects of the volatile components extracted from Pinus densiflora S. and Z., by simultaneous steam distillation and solvent extraction (SDE), were examined on six foodborne bacteria using Bioscreen C (a computer-controlled shake–incubator–reader). The SDE extracts of P. densiflora obtained after 1.5 or 2.0 h at pH 3.6 exhibited a strong growth inhibitory effect on Escherichia coli O157:H7 and their overall antibacterial activities against the various bacteria tested tended to increase with increased extraction time and with a lower extraction pH. The major volatile components of the SDE extracts obtained at pH 3.6 and 1.5 h, as determined by gas chromatography, were α-ocimene (29.3%), sabinene (10.9%), β-myrcene (9.6%), β-caryophyllene (8.0%), β-cadinene (7.3%), α-terpinolene (4.9%), 2-hexanal (4.5%), and β-pinene (4.3%). The addition of 8% or 10% (v/v) of the SDE extracts to culture broth completely inhibited the growths of Bacillus cereus, Salmonella Typhimurium, and Staphylococcus aureus. The intracellular adenosine triphosphate (ATP) concentration of S. Typhimurium treated with P. densiflora extract reduced to 0.165 μm from 0.595 μm, whereas the ATP concentration in culture supernatants was increased to 0.469 μm form 0.065 μm.
The antibacterial activity of water-soluble extract from pine needles of Cedrus deodara (WEC) was evaluated on five food-borne bacteria, and its related mechanism was investigated by transmission electron microscope. In vitro antibacterial assay showed that WEC possesses a remarkable antibacterial activity against tested food-borne bacteria including Escherichia coli, Proteus vulgaris, Staphylococcus aureus, Bacillus subtilis and Bacillus cereus, with the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values in the ranges of 0.78-12.5 mg/ml and 1.56-25mg/ml, respectively. In a food system of fresh-squeezed tomato juice, WEC was observed to possess an effective capacity to control the total counts of viable bacteria. Shikimic acid was isolated from WEC and identified as the main antibacterial compound. All results of our study suggested that WEC might be a new potential source of natural antibacterial agents applicable to food.
Background:
To investigate the volatile compounds and the antibacterial and antioxidant effects of the essential oils of Pinus densiflora needles (EPDN) and Pinus thunbergii needles (EPTN), the volatile compounds of steam-distilled essential oils were analysed by gas chromatography-mass spectrometry. Antibacterial activities were analysed by performing disc-agar diffusion assay and determining the minimum inhibitory concentrations (MICs) of the essential oils. Antioxidant activities were analysed via radical- and nitrite-scavenging activity assays.
Results:
The yields of EPDN and EPTN were 0.304% (v/w) and 0.296% (v/w), respectively. In the antibacterial activity assay, the MICs of EPDN and EPTN for Klebsiella pneumoniae, Shigella flexneri and Proteus vulgaris were < 0.4 mg mL(-1) . In the antioxidant activity assay, the 50% inhibitory concentrations (IC(50) ) of EPDN and EPTN were 120 and 30 µg mL(-1) , respectively. At 1680 µg mL(-1) , both EPDN and EPTN exhibited > 50% nitrite-scavenging activity.
Conclusion:
EPDN can be used as a natural antimicrobial substance.
The antimicrobial effect of linolenic acid with or without monoglyceride (glycerol laurate or glycerol myristate) against six food-borne microorganisms was determined in broth medium. Minimum inhibitory concentration of linolenic acid on Bacillus cereus and Staphylococcus aureus was 20 and 50 ppm, respectively. The growth of B. cereus treated with linolenic acid at 10 ppm with 10 ppm monoglyceride was more inhibitory than that of linolenic acid alone, and the viable cell population was reduced 2-4 log cycles compared to that of the control. When linolenic acid was added at that level, the adenosine triphosphate (ATP) concentration of extracellular fluid was drastically increased compared with that of the control, and the combined effect with monoglyceride was higher than that with linolenic acid alone. However, the intracellular ATP concentration decreased compared with that of the control. From these results, we concluded that linolenic acid has a strong antimicrobial activity against B. cereus and S. aureus, and that linolenic acid combined with monoglyceride showed stronger antimicrobial activity than using linolenic acid alone.
Essential oils and their corresponding hydrosols, obtained after distillation of various scented Pelargonium (Geraniaceae) leaves were assessed for their antimicrobial activity in a model food system. Both the essential oils and hydrosols were used at 1000 ppm in broccoli soup, previously inoculated with Enterobacter aerogenes (at 10(5) cfu g(-1)) and Staphylococcus aureus (at 10(4) cfu g(-1)). The results showed a complete inhibition of S. aureus in the broccoli soup by the essential oils of 'Sweet Mimosa', 'Mabel Grey', P. graveolens, 'Atomic Snowflake', 'Royal Oak', 'Attar of Roses' and a lesser effect by 'Chocolate Peppermint' and 'Clorinda'; the hydrosols, however, had a potentiating effect on the bacterial population in the food. Both extracts showed a complete inhibition of S. aureus in the Maximum Recovery Diluent (MRD). Antibacterial activity against E. aerogenes in the broccoli soup was generally very much reduced: only the essential oil of 'Mabel Grey' showed complete inhibition and virtually no reductions in colonies were seen with the other essential oils; the hydrosols again caused an increase in bacterial colonies. All the essential oils, bar Chocolate Peppermint showed complete inhibition of E. aerogenes in MRD, but the hydrosols showed no effect. The results strongly suggest that the residual hydrosols from distillation of these plant essential oils have no potential as antibacterial agents in foods, in contrast to most of the essential oils, which show potential against some micro-organisms, but only in some food systems. The problem of food component interference and its possible management is discussed.
This article is based on the Shirley Lerner Memorial Lecture given to the Manchester Industrial Relations Society, 18 May 1995. This article draws freely on joint work with my Centre for Economic Performance colleagues Sue Fernie and Stephen Woodland. In particular, it incorporates some material from my joint article with Sue Fernie (1995). Anyone wanting further information on the topics here may wish to consult that article. I acknowledge, with thanks, helpful discussions on this article with Rachel Bailey and Sue Fernie.
Fifty plant essential oils were examined for their antibacterial properties against 25 genera of bacteria. Four concentrations of each oil were tested using an agar diffusion technique. The ten most inhibitory oils were thyme, cinnamon, bay, clove, almond (bitter), lovage, pimento, marjoram, angelica and nutmeg. The most comprehensively inhibitory extracts were angelica (against 25 genera), bay (24), cinnamon (23), clove (23), thyme (23), almond (bitter) (22), marjoram (22), pimento (22), geranium (21) and lovage (20).
The antibacterial effects of four spice components, namely thymol, eugenol, menthol and anethole were tested against Salmonella typhimurium, Staphylococcus aureus and Vibrio parahaemolyticus. Of the spice components tested, eugenol was found to be the most effective followed by thymol, anethole and menthol.