Effects of Essential Oil Vapour Treatment on the Postharvest Disease Control and Different Defence Responses in Two Mango (<Emphasis Type="Italic">Mangifera indica</Emphasis> L.) Cultivars

The purpose of this study was to investigate the inhibitory effect of essential oils (thyme, clove and cinnamon) in vapour phase against the major fungal diseases of mango in vitro and in vivo. Thyme oil vapour (5 μL/Petri plate) completely inhibited the mycelial growth of Colletotrichum gloeosporioides and Lasiodiplodia theobromae under in vitro condition. Thyme oil vapour at 66.7 μL L^?1 significantly reduced artificially inoculated C. gloeosporioides and L. theobromae in mangoes for 4 days. GC/MS analysis revealed thymol, eugenol and benzofuran, 3-methyl as the dominant compounds in thyme, clove and cinnamon oils, respectively. The activities of defence and antioxidant enzymes including peroxidase, chitinase, phenylalanine ammonia-lyase, β-1,3-glucanase, catalase and superoxide dismutase were enhanced by thyme oil (66.7 μL L^?1) treatment and also help to maintain the phenolic content. Hence, postharvest thyme oil vapour treatment may prove to be an alternative means of controlling disease in mangoes.     

The potential application of plant essential oils as natural food preservatives in soft cheese

Investigations were carried out to assess the efficiency of four plant essential oils; bay, clove, cinnamon and thyme as natural food preservatives. The effect of the plant essential oils at concentrations of 0·1, 0·5 and 1% was studied in low-fat and full-fat soft cheese against Listeria monocytogenes and Salmonella enteritidis at 4° and 10°C respectively, over a 14-day period. The composition of the cheese was shown to be an important factor in determining the effectiveness of the plant essential oils. In the low-fat cheese, all four oils at 1% reduced L. monocytogenes to ≤1·0 log₁₀cfu ml⁻¹. In contrast, in the full-fat cheese, oil of clove was the only oil to achieve this reduction. Oil of thyme proved ineffective against S. enteritidis in the full-fat cheese, yet was equally as effective as the other three oils in the low-fat cheese, reducing S. enteritidis to ≤1·0 log₁₀cfu ml⁻¹from day 4 onwards. It is concluded that selected plant essential oils can act as potent inhibitors of L. monocytogenes and S. enteritidis in a food product.     

Chemical composition and antimicrobial activity of the essential oils from four Ruta species growing in Algeria

Antimicrobial properties of plants essential oils have been investigated in order to suggest them as potential tools to overcome the microbial drug resistance and the increasing incidence of food borne diseases problems. The aim of this research is to study the antibacterial and antifungal effects of four traditional plants essential oils, Ruta angustifolia, Ruta chalepensis, Ruta graveolens and Ruta tuberculata, against standard bacterial and fungal strains. The chemical compounds of the oils were examined by GC/MS. Results revealed a powerful antifungal activity against filamentous fungi. Aspergillus fumigatus and Cladosporium herbarum are the most sensitive strains to these oils with MIC values less than 3.5 μg ml⁻¹ for certain oils, reaching 7.8 μg ml⁻¹ for other. GC/MS essay exhibited ketones as the most abundant constituent of these oils except for R. tuberculata essential oil which has a completely different composition, monoterpenes alcohols being the most abundant. These compositions explain their potential antifungal activity.     

Resistance of rubberwood (<Emphasis Type="Italic">Hevea brasiliensis</Emphasis>) treated with chitosan or silane against surface molds

The aim of this study was to determine the efficacy of chitosan and methoxysilane in the prevention of surface mold growth on rubberwood. Three different chitosan samples were tested; C1 (Mw 37 kDa), C2 (Mw 5.4 kDa) and C3 (Mw 3.5 kDa). Radial growth inhibition assay of the chitosan samples was investigated at concentrations ranging from 0.063 to 0.5 %w/v against Aspergillus niger BAM 4 and Penicillium decumbens CBS 121928. Chitosan samples C1 and C3 exhibited strong antifungal activity against both molds. Rubberwood samples were either vacuum or dip treated with varying concentrations of chitosan or silane solution. The content of chitosan in wood showed that after the leaching test, chitosan was well retained in both vacuum and dip treated wood. The concentration of silicon in wood showed similar results. The vacuum treated wood samples with chitosan C1 and C3 at 1 %w/v concentration had strong resistance against A. niger BAM 4. However, dip treated rubberwood samples with 2 %w/v chitosan solutions showed lower resistance against A. niger BAM 4. On the other hand, both vacuum and dip treated rubberwood samples with chitosan had no resistance against P. decumbens CBS 121928. The silane treated wood samples showed no resistance to fungal growth.     

Comparison of releasing bound phenolic acids from wheat bran by fermentation of three Aspergillus species

Wheat bran was fermented at 28 °C for 7 days under 70% humidity by Aspergillus niger, Aspergillus oryzae and Aspergillus awamori. Total phenolic content (TPC) of the unfermented sample was 1531.5 μg g^?1 wheat bran. After the fermentation of Aspergillus awamori, Aspergillus oryzae and Aspergillus niger, TPC reached 5362.1, 7462.6 and 10 707.5 μg g^?1, respectively. The antioxidant activity in the extractions of fermented wheat bran also increased significantly compared with the unfermented sample (P < 0.05). Aspergillus niger showed the greatest capacity to release bound ferulic acid (416.6 μg g^?1). Aspergillus oryzae and Aspergillus awamori had the advantages of releasing more chlorogenic acid (84.0 μg g^?1) and syringic acid (142.3 μg g^?1), respectively. The destructive effect of Aspergillus niger on wheat bran structure was the strongest, followed by that of Aspergillus oryzae. This effect of Aspergillus niger may be due to its higher cellulase, xylanase, arabinofuranosidase and β‐xylosidase activities. Besides, Aspergillus oryzae possessed higher β‐glucosidase activity, and Aspergillus awamori had higher α‐amylase and feruloyl esterase activities. Aspergillus niger may be the best to release bound phenolic acids in the three Aspergillus species. These will provide the helpful information for understanding mechanism of the fermentation by Aspergillus species releasing bound phenolic in wheat bran.     

Determination of Iron in Edible Oil by FAAS After Extraction with a Schiff Base

The synthesis of a new Schiff base has been achieved by condensation reaction of 1,3-diamino-2-propanol and 2-hydroxy-5-methoxy-benzaldehyde in alcoholic media. The Schiff base was characterized by elemental analysis, IR, UV–Visible, ¹H-NMR, and ¹³C-NMR spectroscopy and used for the extraction of iron in liquid edible oils. Iron complex with the Schiff base was investigated in order to determine experimental conditions of the complexation. The extraction of iron from oils to aqueous phase was succeeded by the complexation with the Schiff base. A central composite design was employed in order to optimize the extraction conditions of iron. Optimum conditions for the iron extraction with N,N′-bis(5-methoxy-salicylidene)-2-hydroxy-1,3-propanediamine were found to be 19.3 °C, 2.1 mL g^?1, 10.0 min for the temperature, the ratio of the volume of used Schiff base solution to the amount of oil, and the stirring time, respectively. Method validation was performed with recovery experiments in the analysis of the oil-based metal standard by flame atomic absorption spectrometry, limit of detection, and the relative standard deviation of the proposed method were found to be 0.09 μg g^?1 and 1.04 %, respectively. Additionally, an alternative determination procedure (inductively coupled plasma optical emission spectrometry) was applied for the comparison. This paper describes a simple, cheap, rapid, efficient, sensitive, and accurate alternative analytical method for the iron determination in oils. The proposed method was applied on six different oil samples and the iron concentration was found in the range of 0.38–0.70 μg g^?1.     

Selected plant essential oils and their main active components,a promising approach to inhibit aflatoxigenic fungi and aflatoxin production in food

Recent research has showed that Aspergillus flavus and Aspergillus parasiticus are aflatoxigenic species that can become very competitive in the framework of climate change. Aflatoxins show carcinogenic, mutagenic, immunotoxic and teratogenic effects on human and animals. Effective and sustainable measures to inhibit these species and aflatoxins in food are required. Origanum vulgare and Cinnamomum zeylanicum essential oils (EOs) and their major active constituents, carvacrol and cinnamaldehyde, respectively, were assayed for inhibiting these species and aflatoxin production in maize extract medium under different environmental conditions. Doses of 10–1000 mg l^?1 were assayed and the effective doses for 50 (ED₅₀) and 90% (ED₉₀) growth inhibition were determined. The ED₅₀ of cinnamaldehyde, carvacrol, oregano EO, and cinnamon EO against A. flavus were in the ranges 49–52.6, 98–145, 152–505, 295–560 mg l^?1 and against A. parasiticus in the ranges 46–55.5, 101–175, 260–425 and 490–675 mg l^?1, respectively, depending on environmental conditions. In A. flavus treatments ED₉₀ were in the ranges 89.7–90.5, 770–860 and 820–>1000 mg l^?1 for cinnamaldehyde, carvacrol and cinnamon EO, and in A. parasiticus treatments in the ranges 89–91, 855–>1000 and 900–>1000 mg l^?1, respectively. ED₉₀ values for oregano EO against both species were >1000 mg l^?1. Growth rates of both species were higher at 37 than at 25°C and at 0.99 than at 0.96 a_w. Aflatoxin production was higher at 25 than at 37°C. Stimulation of aflatoxin production was observed at low doses except for cinnamaldehyde treatments. The effectiveness of EOs and their main constituents to inhibit fungal growth and aflatoxin production in contact assays was lower than in vapour phase assays using bioactive EVOH-EO films previously reported.     

Determination of Free Amino Acids in Coated Foods by GC–MS: Optimization of the Extraction Procedure by Using Statistical Design

The development and validation of an extraction procedure for quantification of free amino acids in coated products by MTBSTFA derivatization and GC–MS detection is described. The extraction method entailed the sample homogenization with hydrochloric acid (HCl) by stirring at 40 °C followed by two centrifugation steps. The optimum combination of the extraction variables was achieved by response surface methodology. HCl concentration and volume and stirring time influenced free amino acid extraction yield. The selected optimal extraction conditions were 5 g of sample mixed to 7.5 ml of 0.1 N HCl and stirred during 90 min. Consistency between predicted and experimental values as well as in the quality parameters was observed. The calibration curves were linear within the range 5–100 μg ml^?1 with correlation coefficient values (R ² ) higher than 0.99. Detection and quantification limits of the analytical procedure ranged from 2.10^?5 to 18.10^?2 μg μl^?1 and from 8.10^?5 to 60.10^?2 μg μl^?1, respectively. Precision was 0.20–12.59 % for run-to-run and 3.38–17.60 % for day-to-day. The accuracy is between 82.99 and 115.77 %. Nineteen amino acids were analyzed in frozen-thawed and deep-fried coated products from different origin, with cysteine being the most relevant.     

Effect of oil heating age on colour and dimensional stability of heat treated Pinus radiata

Pinus radiata specimens with moisture content of 8–11% and dimensions of 90×35×200 mm³ were heat-treated in an oil bath of commercial grade raw linseed oil. The effect of oil aging was investigated under treatment condition of 180°C and 3 hours by using oil that had been preheated for 0, 3, 9, 15, 21, and 27 hours, respectively, before the wood treatment. After treatment using oils of varying ages, wood colour change was examined using a Minolta spectrophotometer through CIE 1976 (L ^* a ^* b) system. Water repellent efficiency and anti swelling efficiency in high humidity conditions were measured to determine the stability of the treated wood. The results show that stability of the treated wood was improved significantly compared to the matched untreated wood but the treated wood tended to be darker. Oil viscosity increased with the heating age resulting in slight decrease in weight percentage gain. Water repellent efficiency was decreased with an increase in heating age of oil. However no significant difference in total colour variation and wood stability (anti swelling efficiency) was observed between specimens treated with oils of varying heating age.     

A Modified QuEChERS Sample Preparation Method for the Analysis of 70 Pesticide Residues in Tea Using Gas Chromatography-Tandem Mass Spectrometry

A modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) method for the simultaneous determination of 70 pesticides in tea was developed using gas chromatography-tandem mass spectrometry. Prior to acetonitrile extraction of the target compounds from tea matrix, samples were soaked in distilled water to improve the extraction efficiency. A mixture of adsorbents containing primary–secondary amine, octadecylsilane, graphite carbon black, and multiwalled carbon nanotubes was applied for the cleanup. Additional steps of concentration and solvent exchange were performed to reduce the amount of co-extracts and to decrease the limit of detection of the method. For all pesticides, good linear calibrations with coefficients (R ²) ≥0.99 were obtained at the concentration levels of 10, or 50 to 1,000 μg ml^?1. The limits of quantifications (LOQs) were 5–25 μg ml^?1, respectively. The recovery rates of samples spiked with 20, 100, and 200 μg kg^?1 of analytes ranged from 71 % to 105 %. In addition, the relative standard deviations were lower than 20 %. A total of 331 tea samples were analyzed using this method, and the levels of five pesticide residues in nine tea samples exceeded the strictest maximum residual limits (MRLs).     

Antifungal activity of five different essential oils in vapour phase for the control of Colletotrichum gloeosporioides and Lasiodiplodia theobromae in vitro and on mango

Mango fruit has high commercial value; however, major postharvest losses are encountered throughout the supply chain due to postharvest diseases. These results lead to the search for natural fungicide for postharvest diseases control. The antifungal effects of five essential oils (thyme, clove, cinnamon, anise and vitex) were assessed by disc volatilisation method. Thyme oil vapours at 5 μL per Petriplate, and clove and cinnamon oil at 8 μL per Petriplate showed 100% growth inhibition of mango pathogens in vitro. GC/MS analysis of essential oil showed thymol (23.88), o‐cymol (23.88) and terpinolene (23.88) as the major constituents of thyme oil. Clove and cinnamon oils contain 3‐allyl‐2‐methoxyphenol (37.42%) and benzofuran 3‐methyl (17.97%), respectively. Thyme oil as a fumigant at 66.7 μL L^?1 showed a significant (P < 0.05) inhibition on postharvest pathogens of mango fruits stored at 25 °C for 6 days. Results of our study suggest the possibility of using thyme oil as an alternate natural fungicide to manage postharvest diseases in mango.     

Detoxification of essential oil components (citral and menthol) by Aspergillus niger and Rhizopus stolonifer

The antifungal activity of two essential oil components (citral and menthol) towards Aspergillus niger and Rhizopus stolonifer was studied in shake culture. The activity was high with lower inoculum density (measured in terms of number of spores ml^?1), and it decreased as the inoculum density increased. Citral at 200 μg ml^?1 and menthol at 400 μg mh^?1 were lethal to the spores of A. niger and R. stolonifer, respectively, after a 48-h treatment. At lower concentrations both the compounds were only fungistatic: the growth of the fungi resumed after an initial delay or upon transfer to fresh medium. Periodic estimations of the essential oil components by gas-liquid chromatography showed a continuous decrease in their concentrations in the culture medium, indicating their removal tending to detoxification. The difference in the activity of the two compounds was related to the capacity of the fungi to detoxify the compounds.     

Assessment of Thymus vulgaris L. essential oil as a safe botanical preservative against post harvest fungal infestation of food commodities

A total of 14 odoriferous angiospermic essential oils were tested against the toxigenic strain of Aspergillus flavus. The essential oil of Thymus vulgaris L. showed highest antifungal efficacy. The thyme oil absolutely inhibited the mycelial growth of A. flavus at 0.7 μl ml^− 1 and exhibited a broad fungitoxic spectrum against eight different food contaminating fungi viz. Fusarium oxysporum, Cladosporium herbarum, Curvularia lunata, Aspergillus terreus, Aspergillus niger, Aspergillus fumigatus, Alternaria alternata and Botryodiploidia theobromae. The oil also showed significant antiaflatoxigenic efficacy as it completely arrested the aflatoxin B₁ production at 0.6 μl ml^− 1. Thyme oil as fungitoxicant was also found superior over most of the prevalent synthetic fungicides. The LC₅₀ of thyme oil against mice was recorded as 7142.85 μl kg^− 1 body weight indicating its non-mammalian toxicity and strengthening its safe exploitation as preservative for stored food commodities. The findings recommend the thyme oil as potential botanical preservative in eco-friendly control of biodeterioration of food commodities during storage.

Industrial relevance

The thyme essential oil may be recommended for large scale application as a plant based preservative for stored food items because of its strong antifungal as well as antiaflatoxigenic efficacy. Because of broad antimicrobial spectrum, more efficacy over prevalent synthetic preservatives as well as non-mammalian toxicity, the thyme essential oil may be formulated as a safe and economical plant based preservative against post harvest fungal infestation and aflatoxin contamination of food commodities.     

Ultrasonic-Assisted Extraction of α-Tocopherol Antioxidants from the Fronds of Elaeis guineensis Jacq.: Optimization, Kinetics, and Thermodynamic Studies

This article presents the first report on the extraction and quantification of α-tocopherol from the fronds of the oil palm (Elaeis guineensis Jacq). In this study, the optimization, kinetic, and thermodynamic data of α-tocopherol extraction by sonication are presented. Response surface methodology coupled with central composite design was used to optimize the experimental conditions for α-tocopherol extraction. Three independent variables, namely sample/solvent ratio (1:20–1:40 g/ml), extraction temperature (30–50 °C), and extraction time (20–50 min) were studied. For optimum conditions of 39.31 °C (~40 °C), 50 min, and 1:23.63 g/mL (~1:20 g/mL), total tocols and α-tocopherol optimal concentrations were 346.49 μg/g oil palm fronds (OPFs) by dry weight (DW) and 28.41 μg/g OPF DW, respectively. The effects of extraction temperature and tocol concentrations on the extraction kinetics and thermodynamic parameters were also studied. From the mass transfer rate equation, the kinetic and thermodynamic data obtained from the ultrasonic-assisted extraction (UAE) of α-tocopherol from OPF were activation energy, E _a (104.6 kJ mol^?1), UAE rate constant, k (6.886?×?10^?3 min^?1), ΔH (+0.818 kJ mol^?1), ΔS (+27.22 J mol^?1 K^?1), and ΔG (?8.52 J mol^?1). According to this study, the UAE of α-tocopherol from OPF is endothermic, irreversible, and spontaneous.     

Post-harvest practices linked with ochratoxin A contamination of coffee in three provinces of Cordillera Administrative Region,Philippines

One of the emerging concerns in the Cordillera Administrative Region, Philippines is ochratoxin A (OTA) contamination in coffee. During 2015 to 2016, a total of 51 Arabica (Coffea arabica) coffee samples from Benguet province and 71 Robusta (Coffea canephora var. Robusta) coffee samples from the provinces of Ifugao and Kalinga were analysed for OTA contamination. The OTA-producing fungal contaminants during drying and storage of Arabica and Robusta coffee were Aspergillus niger and Aspergillus ochraceus. Ochratoxin A was more commonly detected in Robusta coffee (36.6%) than in Arabica coffee (21.6%). Among the contaminated samples, Robusta coffee cherries in the drying yard had the highest mean OTA level (120.2 μg kg^?1, n = 10) while roasted Robusta coffee beans had the lowest mean level (4.8 μg kg^?1, n = 9). The onset of contamination of Arabica coffee occurred during storage, with a mean OTA level of 46.7 μg kg^?1 (n = 9). Roasted coffee had lower OTA content although five samples had levels >5.0 μg kg^?1. Pearson Chi-square analysis (χ²) and Fisher’s exact test revealed that several post-harvest practices involving non-removal of the husk or hull and mixing of defective coffee were significantly associated with the occurrence of OTA during drying and storage (p < 0.05). No significant associations, however, were identified during roasting. This study suggests that the post-harvest practices in Cordillera Administrative Region should focus on the removal of defective coffee in all stages of post-harvest and rapid reduction of moisture content particularly during drying.     

Physicochemical Characterization of Lemongrass Essential Oil–Alginate Nanoemulsions: Effect of Ultrasound Processing Parameters

The formation of lemongrass oil (1 %?v/v) nanoemulsions in aqueous sodium alginate solution (1 %?w/v) containing Tween 80 (1 %?v/v) as nonionic surfactant was studied in terms of droplet size, electrical charge, viscosity, and whiteness index considering different ultrasonication times (0, 30, 60, 120, and 180 s) and amplitudes (30, 60, and 100 μm). The droplet size and size distribution of the emulsions decreased at increasing treatment time and amplitude. The minimum average droplet size observed in nanoemulsions was 4.31?±?0.18 nm with a narrow size distribution. The interface electrical charge of the coarse emulsion was ?18.0?±?2.9 mV, whereas in ultrasonicated nanoemulsions, it diminished up to ?55.8?±?6.4 mV when the sonication time was extended for 180 s. The viscosity of nanoemulsions also decreased at increasing treatment time and amplitude. Moreover, nanoemulsions became translucent after sonicating for 180 s at 30, 60, or 100 μm with whiteness indices of 28.61?±?0.17, 27.93?±?0.21, and 27.86?±?0.33, respectively. Therefore, it can be stated that ultrasound processing might be a feasible technology to produce highly translucent lemongrass oil–alginate nanoemulsions, with extremely small droplet sizes and high stability to be used as delivery systems of essential oils in food products. However, it is necessary to investigate the effect of ultrasound processing parameters on the antimicrobial potential of essential oils incorporated to nanoemulsions.     

Occurrence of 3-MCPD and glycidyl esters in edible oils in the United States

Fatty acid esters of 3-monochloropropanediol (3-MCPD) and glycidol are processing contaminants found in a wide range of edible oils. While both 3 MCPD and glycidol have toxicological properties that at present has concerns for food safety, the published occurrence data are limited. Occurrence information is presented for the concentrations of 3-MCPD and glycidyl esters in 116 retail and/or industrial edible oils and fats using LC-MS/MS analysis of intact esters. The concentrations for bound 3-MCPD ranged from below the limit of quantitation (<LOQ) to 0.09 mg kg^?1 (ppm) in 22 unrefined oils and from 0.005 to 7.2 mg kg^?1 (ppm) in 94 refined oils. The concentrations for bound glycidol ranged from <LOQ to 0.03 mg kg^?1 (ppm) in unrefined oil samples and from <LOQ to 10.5 mg kg^?1 (ppm) in processed oil samples. The highest concentrations for both 3-MCPD and glycidol were seen in refined palm oil and palm olein samples. Palm olein samples also contained a higher percentage of 3-MCPD in mono-ester form than any other type of oil.     

Inactivation of Escherichia coli O157:H7 by cinnamic aldehyde purified from Cinnamomum cassia shoot

Escherichia coli O157:H7 is a pathogen, which causes the hemorrhagic colitis, hemolytic uremic syndrome and thrombotic thrombocytopenic purpura in humans. Control of the bacterial cells in foods is an important factor to reduce outbreaks of the foodborne diseases. In this study, cinnamic aldehyde possessing antimicrobial activity against the bacterial cells was purified from the extract of cinnamon (Cinnamomum cassia Blume) shoot by sequential fractionation with various solvents and silica gel column chromatography. When E. coli O157:H7 cells were incubated at 37°C for 12 h in the presence of 500 μg ml⁻¹ of the purified from cinnamic aldehyde, the viable counts decreased dramatically (from 4.9×10⁶ to 1.0×10² cfu ml⁻¹). In the presence of 1000 μg ml⁻¹ of the substance, most of the cells were killed after 2 h of incubation suggesting that the antimicrobial activity of cinnamic aldehyde is bacteriocidal in E. coli. Scanning electron microscopic observations revealed that the bacterial cells treated with the cinnamic aldehyde suffered from severe damages in their surface structure. Minimal inhibitory concentration of the cinnamic aldehyde was determined to be 250 μg ml⁻¹ against E. coli strains O157:H7 and O26 or 500 μg ml⁻¹ against strains ATCC11105 and O111.     

Unravelling a vicious circle: animal feed marketed in Costa Rica contains irregular concentrations of tetracyclines and abundant oxytetracycline-resistant Gram-positive bacteria

Diverse tetracyclines are used to prevent and control bacterial infections in livestock and farmed fish. These drugs are administered through the diet, but farmers seldom check whether feed contains antibiotic-resistant bacteria that may colonise their crops or transfer their resistance traits to species of veterinary relevance. To examine whether antibiotic dosage defines the abundance of antibiotic-resistant bacteria in animal feed, we determined the concentration of parental compounds and epimers of oxytetracycline (OTC), doxycycline, tetracycline and chlortetracycline, as well as the abundance and resistance level of OTC-resistant bacteria in samples of fish (n = 21), poultry (n = 21), swine (n = 21), and shrimp feed (n = 21) marketed in Costa Rica. Fish feed contained the highest amounts of tetracyclines (119–8365 mg kg^?1) and the largest proportion of bacteria resistant to 10 μg ml^?1 (1.8–92.4%) or 100 μg ml^?1 of OTC (12.5–63.8%). Poultry (78–438 mg kg^?1) and swine (41–1076 mg kg^?1) feed had intermediate concentrations of tetracyclines and OTC-resistant bacteria (0.2–66% and 0.3–49%, respectively), whereas shrimp feed showed the lowest amounts of tetracyclines (21.5–50.3 mg kg^?1), no OTC and no culturable OTC-resistant bacteria. In line with these results, the MIC₅₀ of OTC for 150 isolates from fish and poultry feed was > 256 µg ml^?1, while that of 150 bacteria isolated from swine feed was 192 µg ml^?1. Phenotypic tests, fatty acid profiles and proteotypic analyses by matrix-assisted laser desorption/ionisation-time of flight mass-spectroscopy revealed that most OTC-resistant isolates were Gram-positive bacteria of low G+C% content from the genera Staphylococcus and Bacillus. Clear correlations between OTC dosage and feed colonisation with OTC-resistant bacteria were seen in medicated feed for fish (r = 0.179–0.651). Nonetheless, some unmedicated feed for fish, swine and poultry contained large populations of OTC-resistant bacteria, suggesting that raw materials and manufacturing processes may also influence carriage of OTC-resistant bacteria in animal feed.     

Tocopherols and Tocotrienols: an Adapted Methodology by UHPLC/MS Without Sample Pretreatment Steps

Ultra high-performance liquid chromatography-electrospray ionization-mass spectrometry (UHPLC/ESI/MS) methodology was adapted for identification and quantification of tocopherols and tocotrienols in vegetable oils with no derivatization or sample preparation steps. The UHPLC analysis was performed using a C18 column and mobile phase composed of methanol: water: ammonium hydroxide (99:1:0.1 v/v/v) and isopropanol. A single mass spectrometer with electrospray on negative mode was used as a detector for tocopherols and tocotrienols. The samples were diluted in isopropanol. The limit of quantification for tocopherols was 0.006 μg mL^?1, and the linear range was 0.006 to 0.01 μg mL^?1; for tocotrienols, the limit of quantification was 0.002 μg mL^?1, and the linear range of analysis was 0.002 to 0.003 μg mL^?1. The correlation coefficients were higher than 0.99, indicating that the method has suitable linearity. The methodology has proven to be precise, reproducible, and robust for the parameters studied.