In vitro effects of citrus oils against Mycobacterium tuberculosis and non-tuberculous Mycobacteria of clinical importance

We evaluated the in vitro activity of citrus oils against Mycobacterium tuberculosis and other non-tuberculous Mycobacterium species. Citrus essential oils were tested against a variety of Mycobacterium species and strains using the BACTEC radiometric growth system. Cold pressed terpeneless Valencia oil (CPT) was further tested using the Wayne model of in vitro latency.

Exposure of M. tuberculosis and M. bovis BCG to 0.025 % cold pressed terpeneless Valencia orange oil (CPT) resulted in a 3-log decrease in viable counts versus corresponding controls. Inhibition of various clinical isolates of the M. avium complex and M. abscessus ranged from 2.5 to 5.2-logs. Some species/strains were completely inhibited in the presence of CPT including one isolate each of the following: the M. avium complex, M. chelonae and M. avium subsp. paratuberculosis. CPT also inhibited the growth of BCG more than 99 % in an in vitro model of latency which mimics anaerobic dormancy thought to occur in vivo. The activity of CPT against drug-resistant strains of the M. avium complex and M. abscessus suggest that the mechanism of action for CPT is different than that of currently available drugs. Inhibition of latently adapted bacilli offers promise for treatment of latent infections of MTB. These results suggest that the antimycobacterial properties of CPT warrant further study to elucidate the specific mechanism of action and clarify the spectrum of activity.     

Ciprofloxacin-sensitive and ciprofloxacin-resistant Campylobacter jejuni are equally susceptible to natural orange oil-based antimicrobials

A total of 10 ciprofloxacin-sensitive (ciprofloxacin minimum inhibitory concentration, MIC < 0.5 μ g/ml) and 10 ciprofloxacin-resistant (MIC 16 to 32 μ g/ml) presumptive C. jejuni were further characterized and evaluated for their inhibition by natural orange oil fractions. Partial species identification was performed by using a hippuricase gene-based polymerase chain reaction (PCR) assay. One of the isolates appeared to be atypical and failed to hydrolyze hippurate. Of the ciprofloxacin-resistant C. jejuni isolates tested, six were found to have their quinolone resistance determined by a C → T mutation in codon 86 of gyrA. Both groups of ciprofloxacin-sensitive and –resistant C. jejuni isolates were most susceptible to cold-pressed terpeneless Valencia orange oil (C4) which yielded inhibition zones from 44.0 ± 1.4 to 80 ± 0.0 mm. Less inhibitory responses were recorded for 5-fold concentrated Valencia orange oil (C3) and distilled d-limonene (C7) which exerted similar effects on both ciprofloxacin-sensitive and –resistant C. jejuni isolates. In general, ciprofloxacin-resistant and –sensitive C. jejuni isolates were equally susceptible to the respective orange oil fractions.     

Toxicity of twenty-two plant essential oils against pathogenic bacteria of vegetables and mushrooms

Toxicity of twenty-two essential oils to three bacterial pathogens in different horticultural systems: Xanthomonas campestris pv. phaseoli (causing blight of bean), Clavibacter michiganensis subsp. michiganensis (bacterial wilt and canker of tomato), and Pseudomonas tolaasii (causal agent of bacterial brown blotch on cultivated mushrooms) was tested. Control of bacterial diseases is very difficult due to antibiotic resistance and ineffectiveness of chemical products, to that essential oils offer a promising alternative. Minimal inhibitory and bactericidal concentrations are determined by applying a single drop of oil onto the inner side of each plate cover in macrodilution assays. Among all tested substances, the strongest and broadest activity was shown by the oils of wintergreen (Gaultheria procumbens), oregano (Origanum vulgare), and lemongrass (Cymbopogon flexuosus. Carvacrol (64.0–75.8%) was the dominant component of oregano oils, while geranial (40.7%) and neral (26.7%) were the major constituents of lemongrass oil. Xanthomonas campestris pv. phaseoli was the most sensitive to plant essential oils, being susceptible to 19 oils, while 11 oils were bactericidal to the pathogen. Sixteen oils inhibited the growth of Clavibacter michiganensis subsp. michiganensis and seven oils showed bactericidal effects to the pathogen. The least sensitive species was Pseudomonas tolaasii as five oils inhibited bacterial growth and two oils were bactericidal. Wintergreen, oregano, and lemongrass oils should be formulated as potential biochemical bactericides against different horticultural pathogens.     

Formulas of components of citronella oil against mosquitoes (Aedes aegypti)

The mosquito Aedes aegypti is an epidemic vector of several diseases such as dengue fever and yellow fever. Several pesticides are used to control the mosquito population. Because of their frequent use, some mosquitoes have developed resistance. In this study, we used the Y-tube olfactometer to test essential oils of Cymbopogon species and screened specific formulas of components as repellents against Ae. aegypti. At 400 μL, the extracted oil of citronella grass (Cymbopogon nardus) and myrcene produced a low-active response by inhibiting mosquito host-seeking activity. Citronella grass, lemon grass (Cymbopogon citratus), citral and myrcene also produced a low-treatment response to repellents, for more potential to affect host-seeking behavior. Furthermore, the mixture of citral, myrcene, and citronellal oil (C:M:Ci = 6:4:1) greatly affected and inhibited host-seeking behavior (76% active response; 26% treatment response with 40 μL; 42.5%, 18% with 400 μL; and 19%, 23% with 1000 μL). As compared with the result for N,N-diethyl-3-methylbenzamide (DEET; 44%, 22% with 400 μL), adjusting the composition formulas of citronella oil had a synergistic effect, for more effective repellent against Ae. aegypti.     

Antibacterial activity against Clostridium genus and antiradical activity of the essential oils from different origin

In the present study, the antimicrobial and antiradical activities of 15 essential oils were investigated. The antimicrobial activities were determined by using agar disc diffusion and broth microdilution methods against Clostridium genus and antioxidant properties of essential oils by testing their scavenging effect on DPPH radicals activities. We determined the antibacterial activity of Clostridium butyricum, Clostridium hystoliticum, Clostridium intestinale, Clostridium perfringens and Clostridium ramosum. We obtained the original commercial essential oils samples of Lavandula angustifolia, Carum carvi, Pinus montana, Mentha piperita, Foeniculum vulgare Mill., Pinus sylvestris, Satureia montana, Origanum vulgare L. (2 samples), Pimpinella anisum, Rosmarinus officinalis L., Salvia officinalis L., Abies alba Mill., Chamomilla recutita L. Rausch and Thymus vulgaris L. produced in Slovakia (Calendula a.s., Nova Lubovna, Slovakia). The results of the disk diffusion method showed very high essential oils activity against all tested strains of microorganisms. The best antimicrobial activity against C. butyricum was found at Pimpinella anisum, against C. hystoliticum was found at Pinus sylvestris, against C. intestinale was found at Satureia hortensis L., against C. perfringens was found at Origanum vulgare L. and against C. ramosum was found at Pinus sylvestris. The results of broth microdilution assay showed that none of the essential oils was active against C. hystoliticum. The best antimicrobial activity against C. butyricum was found at Abies alba Mill., against C. intestinale was found at Abies alba Mill., against C. perfringens was found at Satureia montana and against C. ramosum was found at Abius alba and Carum carvi. Antioxidant DPPH radical scavenging activity was determined at several solutions of oil samples (50 μL.mL^?1–0.39 μL.mL^?1) and the best scavenging effect for the highest concentration (50 μL.mL^?1) was observed. The antioxidant properties were different in particular plant species. The highest% of inhibition after 30 min. of reaction was observed at Origanum vulgare (93%), Satureia montana (90.66%) and Lavandula augustifolia (90.22%).     

Importance of cholinesterase kinetic parameters in environmental monitoring using estuarine fish

The aim of the present study was to determine the kinetic parameters and cholinesterase (ChE) activity in two teleost fish, the white mouth croaker Micropogonias furnieri (Scianidae) and the Madamango sea catfish Cathorops spixii (Ariidae), to verify their suitability as sentinel species of aquatic pollution by anticholinergic compounds. Individuals of each species were captured in one reference and one polluted site in the Southern Brazilian coast. Brain tissue was used as enzyme source. Inhibition kinetic parameters indicated that ChE from C. spixii collected at the reference site showed more affinity (Ka) for eserine that from those collected at the polluted site. The opposite was observed for the carbamylation constants (Kc). Thus, similar inhibition constants (Ki) were observed. M. furnieri brain showed an extremely low sensitivity to in vitro inhibition by eserine, indicating that it is not a suitable biomarker to be employed in environmental monitoring of anticholinergic compounds. Results from the present study also point to the need for considering kinetic analysis when cholinesterase activity is employed as a biomarker.     

The in vitro effect of selected essential oils on the growth and mycotoxin production of Aspergillus species

The aim of the present study was to assess the antifungal and anti-toxinogenic activity of 15 essential oils (EOs) against three fungi of the genus Aspergillus (A. parasiticus KMi-227-LR, A. parasiticus KMi-220-LR and A. flavus KMi-202-LR). The minimum inhibitory doses (MIDs) of the tested essential oils and their antifungal activity were determined using the micro-atmosphere method. The original commercial essential oil samples of Jasminum officinale L., Thymus vulgaris L., Syzygium aromaticum (L.) Merrill &; Perry, Rosmarinus officinalis L., Ocimum basilicum L., Eucalyptus globulus Labill., Salvia officinalis L., Citrus limon (L.) Burm, Origanum vulgare L., Lavandula angustifolia Mill., Carum carvi L., Citrus sinensis (L.) Osbeck., Zingiber officinalis Rosc., Mentha piperita L. and Cinnamomum zeylanicum Nees. (C. verum J.S.Presl.) were produced in Slovakia (Calendula a.s., Nová ?ubovňa, Slovakia). All essential oils exhibited activity against all tested strains of fungi. After 14 days of incubation, A. flavus (KMi-202-LR) showed the highest susceptibility with a growth inhibition percentage (GIP) of 18.70% to C. limon and 5.92% to C. sinensis, while A. parasiticus (KMi-220-LR) exhibited a GIP of 20.56% to J. officinale. The minimum inhibitory doses (MIDs) of EOs with the most significant activity were recorded. The best antifungal activity, using the micro-atmosphere method was found in S. aromaticum with an MID of 62.5 μL L^?1 air, T. vulgaris (MID of 62.5 μL L^?1 air) and O. vulgare (MID of 31.5 μL L^?1 air) against all tested strains. Mycotoxin production of the tested strains was evaluated by the thin layer chromatography (TLC) method. Mycotoxin production of AFB₁ and AFG₁ was inhibited following all treatments with C. carvi, R. officinale and S. officinale, Eucalyptus globulus L. and O. basilicum L. Essential oils exhibited a potential inhibition activity against toxic fungi, although, these affected only the production of AFB₁.     

Effect of decabromodiphenyl ether and antimony trioxide on controlled pyrolysis of high-impact polystyrene mixed with polyolefins

The controlled pyrolysis of polyethylene/polypropylene/polystyrene mixed with brominated high-impact polystyrene containing decabromodiphenyl ether as a brominated flame-retardant with antimony trioxide as a synergist was performed. The effect of decabromodiphenyl ether and antimony trioxide on the formation of its congeners and their effect on distribution of pyrolysis products were investigated. The controlled pyrolysis significantly affected the decomposition behavior and the formation of products. Analysis with gas chromatograph with electron capture detector confirmed that the bromine content was rich in step 1 (oil 1) liquid products leaving less bromine content in the step 2 (oil 2) liquid products. In the presence of antimony containing samples, the major portion of bromine was observed in the form of antimony bromide and no flame-retardant species were found in oil 1. In the presence of synergist, the step 1 and step 2 oils contain both light and heavy compounds. In the absence of synergist, the heavy compounds in step 1 oil and light compounds in step 2 oils were observed. The presence of antimony bromide was confirmed in the step 1 oils but not in step 2 oils.     

Lethal and sublethal effects of four essential oils on the egg parasitoids Trissolcus basalis

The essential oils from leaves of Schinus molle var. areira, Aloysia citriodora, Origanum vulgare and Thymus vulgaris have showed potential as phytoinsecticides against the green stink bug, Nezara viridula. In this work were evaluated their toxicological and behavioral effects on the parasitoid Trissolcus basalis, a biological control agent of this pest insect. Essential oils were obtained via hydrodestillation from fresh leaves. Insecticide activity in T. basalis females was evaluated in direct contact and fumigation bioassays. Behavioral effects were evaluated in olfactometer bioassays. To evaluate the residual toxicity, females of the parasitoids were exposed to oil residues; in these insects, the sublethal effects were evaluated (potential parasitism and survivorship of immature stages). The essential oils from O. vulgare and T. vulgaris proved to be highly selective when used as fumigant and did not change parasitoid behavior. After one week, the residues of these oils were harmless and did not show sublethal effects against T. basalis. According with these results, essential oils have potential applications for the integrated management of N. viridula.     

Fate of bromine in pyrolysis of printed circuit board wastes

Behavior of Br in pyrolysis of the printed circuit board waste with valuable copper and oil recycling has been studied in the present work. Experimentally, pyrolysis of the printed circuit board waste generated approximately 40.6% of oils, 24.9% of noncondensible gases and 34.5% of solid residues that enriched in copper (90-95%). The cuts of the oils produced from pyrolysis of the printed circuit board waste into weighted boiling fraction were primarily light naphtha and heavy gas oil. Approximately 72.3% of total Br in the printed circuit board waste were found in product gas mainly as HBr and bromobenzene. However, by extended X-ray absorption fine structural (EXAFS) spectroscopy, Cu-O and Cu-(O)-Cu species with bond distance of 1.87 and 2.95 A, respectively, were observed in the solid residues. Essentially, no Cu-Br species was found.     

Detailed characterization of polar compounds of residual oil in contaminated soil revealed by Fourier transform ion cyclotron resonance mass spectrometry

Effects of remediation technologies on polar compounds of crude oil in contaminated soils have not been well understood when compared to hydrocarbons. In this study, ultrahigh resolution Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) was used to characterize the changes in NSO polar compounds of crude oil and residual oil after long-term natural attenuation, biostimulation and subsequent ozonation following biostimulation of contaminated soils. N₁ and O₁ species, which were abundant in the crude oil, were selectively biodegraded, and species with higher double bond equivalent values and smaller carbon numbers appeared to be more resistant to microbial alteration. O₂-O₆ species were enriched by biodegradation and contained a large number of compounds with a high degree of unsaturation. Ozone could react with a variety of polar compounds in residual oil after biodegradation and showed high reactivity with polar species containing aromatic or multi-aliphatic rings, including the residual N₁ and O₁ species, naphthenic acids and unsaturated O₃-O₆ compounds. Fatty acids and O₃-O₈ species dominated by saturated alkyl compounds were resistant to ozonation or the primarily incomplete ozonation products. Principal component analysis of identified peaks in the FT-ICR MS spectra provided a comprehensive overview of the complex samples at the molecular level and the results were consistent with the detailed analysis. Taken together, these results showed the high complexity of polar compounds in residual oils after biodegradation or ozonation in contaminated soil and would contribute to a better understanding of bioremediation and ozonation processes.     

Pyrolysis-GC-MS analysis of crude and heavy fuel oil asphaltenes for application in oil fingerprinting

A Pyrolysis – Gas Chromatography – Mass Spectrometry (Py-GC-MS) method was developed for the analysis and profiling of crude and heavy fuel oil asphaltenes, for application in oil fingerprinting. Asphaltenes were precipitated from ten geographically different oils using n-pentane, and analysed by Py-GC-MS. Alkane profiles and sulphur/aromatic profiles were used to compare the oils, and to correctly differentiate oils from different geographical regions. Py-GC-MS could not differentiate a weathered oil sample and a fresh oil sample from the same source. The results of this study support the findings from a previously developed FTIR method for asphaltene profiling.     

Chemical composition and antimicrobial activities of the essential oil from the seeds of Enterolobium contortisiliquum (leguminosae)

Seeds of Enterolobium contortisiliquum were subjected to steam distillation to obtain a light yellow essential oil in a yield of 3 ml/kg of seeds. The major components of the oil were identified using gas chromatography/mass spectrometry (GC-MS) and were furfural, limonene, linalool, estragole, carvone, and apiole with carvone representing more than 50% of the total composition. Antimicrobial activities of the essential oil were determined against four species of gram positive bacteria (Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, Micrococcus luteus) and two gram negative bacteria (Klebsiella pneumoniae, Serratia Marcescencs). The essential oil inhibited the growth of all tested bacteria but was most effective against the gram positive bacteria. Chemicals that are responsible for the antibacterial effect of the essential oil were determined using the bio-autography thin layer chromatography (TLC) technique. The active compounds responsible for the activity were found to be carvone and estragole.     

The effects of oil spill and clean-up on dominant US Gulf coast marsh macrophytes: a review

The objective of this review was to synthesize existing information regarding the effects of petroleum hydrocarbons on marsh macrophytes in a manner that will help guide research and improve spill-response efficiency. Petroleum hydrocarbons affect plants chemically and physically. Although plants sometime survive fouling by producing new leaves, even relatively non-toxic oils can stress or kill plants if oil physically prevents plant gas-exchange. Plant sensitivity to fouling varies among species and among populations within a species, age of the plant, and season of spill. Physical disturbance and compaction of vegetation and soil associated with clean-up activities following an oil spill appear to have detrimental effects on the US Gulf coast marshes. Other techniques, including the use of chemicals such as cleaners or bioremediation, may be necessary to address the problem. Clean-up may also be beneficial when timely removal prevents oil from migrating to more sensitive habitats.     

脂肪酰基氨基酸对矿物润滑油生物降解性的影响研究

通过生物降解试验研究了N-月桂酰基谷氨酸、N-月桂酰基甘氨酸、N-月桂酰基丙氨酸和N-油酰基甘氨酸4种脂肪酰基氨基酸对HVI 350矿物润滑油生物降解性的影响，采用高倍电子显微镜分析了生物降解过程中微生物数量和形态变化。结果表明，在矿物润滑油中添加少量脂肪酰基氨基酸后，矿物润滑油的生物降解性能明显改善，且生物降解过程中微生物数量增多、形态发生变化，这可能是脂肪酰基氨基酸增加了微生物的营养，且具有表面活性，有利于细胞的吸收，从而促进了矿物润滑油生物降解。     

Chemical composition and antimicrobial activities of the essential oil from the seeds of Enterolobium contortisiliquum (leguminosae)

Seeds of Enterolobium contortisiliquum were subjected to steam distillation to obtain a light yellow essential oil in a yield of 3 ml/kg of seeds. The major components of the oil were identified using gas chromatography/mass spectrometry (GC-MS) and were furfural, limonene, linalool, estragole, carvone, and apiole with carvone representing more than 50% of the total composition. Antimicrobial activities of the essential oil were determined against four species of gram positive bacteria (Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, Micrococcus luteus) and two gram negative bacteria (Klebsiella pneumoniae, Serratia Marcescencs). The essential oil inhibited the growth of all tested bacteria but was most effective against the gram positive bacteria. Chemicals that are responsible for the antibacterial effect of the essential oil were determined using the bio-autography thin layer chromatography (TLC) technique. The active compounds responsible for the activity were found to be carvone and estragole.     

Pressurised liquid-liquid extraction. An approach to the removal of inorganic non-metal species from used industrial oils

Modified pressurised hot water is used for the development of a high pressure liquid-liquid extraction method for the decontamination of used industrial oils from inorganic non-metal species (chlorine, fluorine and sulphur). The oils were subjected to dynamic extraction with water modified with 5% v/v HNO3 at 200 degrees C as extractant. Under these working conditions the analytes were transferred to the aqueous phase. Spontaneous separation of the two immiscible liquid phases (the used oil and extract) takes place in the collection flask after extraction. The treated and untreated oil samples were oxidised and the chloride, fluoride and sulphate thus formed were determined by ion-chromatography. The method was applied to four oil samples from different locations in Spain. A residence time of approximately =10 min provided oil samples from which 88.3%, 89.4% and 89.4% of chloride, fluoride and sulphate, respectively, have been removed with respect to the initial concentration of each analyte in the oil. The repeatability, expressed as relative standard deviation (RSD), was of 11.9%, 13.7% and 7.2% for Cl(-), F(-) and SO4(2-), respectively; whilst the within-laboratory reproducibility yielded RSDs of 6.2%, 7.9% and 6.2% for the same analytes. The proposed approach has proved to be efficient, simple, easily transferable to industrial scale, cheap, fast and environmentally friendly.     

Measurement of fluorescence decay of crude oil: a potential technique to identify oil slicks

The single photon counting technique has been used to measure the decay time for several Kuwaiti crude and refined oils. Fluorescent characteristics of two different bunker oils from widely separate geographic areas have been studied. Laser induced fluorescent data for the crude oil samples are compared with the decay time results to evaluate the potential of these techniques for the identification of oil sticks in the marine environment. Response of the oil film thickness to various excitation wavelengths from a CW Ar ion laser and a pulsed xenon lamp has been investigated.     

Toxic risk associated with sporadic occurrences of Microcystis aeruginosa blooms from tidal rivers in marine and estuarine ecosystems and its impact on Artemia franciscana nauplii populations

Microcystis aeruginosa is a species of freshwater cyanobacteria which can form harmful algal blooms in freshwater water bodies worldwide. However, in spite its sporadic occurrences for short periods of time in estuarine waters, their influence on zooplankton populations present in these ecosystems has not been extensively studied. In this work, Artemia franciscana was used as test organism model, studying mortality against several strains of M. aeruginosa with different degrees of toxigenicity, measuring whole-live cells and homogenate extracts. Results were compared with microcystin-LR equivalent content, measured by immunoassay. The results show that there were no significant differences between both exposure models (whole cells and extracts), and there are significant differences respect to the toxigenicity of cyanobacterial blooms depending of the M. aerugionosa strain involved in the process. Analysis of microcystin-LR equivalent concentration test immediately below the lowest significant concentration in all M. aerugionosa strains was used to determine the potential risk associated with the cell densities during a bloom. Comparison among the selected M. aerugionsa strains show that these factors have influence in the results obtained, and thus, several differences have been evidenced depending of the microcystin-LR equivalent production and the strain type involved.     

Multidimensional gas chromatographic enantiomer quantification of some chlorinated xenobiotics in cod liver and fish oils.

Four chiral HRGC column systems for the separation of selected organochlor pesticides and photoconversion products (heptachlor, cis- and trans-chlordane, o,p'-DDT, o,p'-DDD, and alpha-HCH) were tested. Cod hver oil and fish oil samples from different countries were comparatively analyzed. Of all these columns, the fourth was the most suitable one for the analysis of the selected compounds. The enantiomer ratios [ER] of cis-chlordane, trans-chlordane, photodieldrin, alpha-HCH, and o,p'-DDD in fish oils are nearly 1.0, while the ERs of the same substances are significantly different from 1.0 in cod liver oils. Contrary to that, the ER values of o,p'-DDT are remarkable different from 1 in the cod liver oils as well as in the fish oils.