Studying different dimensions of amylose–long chain fatty acid complexes: Molecular, nano and micro level characteristics

Recently amylose inclusion complexes, or V-amylose, have drawn much attention as a possible vehicle for the nanoencapsulation of unsaturated fatty acids. This study aimed to study three different structural strata of V-amylose, the molecular attributes using XRD, DSC and ¹³C CP/MAS NMR and the nanostructures using SAXS. Using these methods it was noted that decreased degree of fatty acid unsaturation induces the formation of more organized and well defined structures. Specifically, calculations based on SAXS data show that regardless of the crystallization temperature saturated SA yields the highest values for parameters like average crystalline lamellar thickness (φ = 0.26) and characteristic particle dimension (R_g = 9.6). SEM shows this trend extends even into the microscopic level. Overall, this study shows that in the case of long chain fatty acids, increased fatty acid unsaturation impairs the structure of amylose inclusion complexes.     

Effects of long chain fatty acid unsaturation on the structure and controlled release properties of amylose complexes

The addition of functional long chain (LC) unsaturated fatty acids to foods is often hampered by their instability. This study aimed to probe the possibility of using molecular inclusion complexes formed by amylose, termed V-amylose, as a possible solution for the delivery of long chain (LC) fatty acids to the consumer. X-ray diffraction, light scattering and atomic force microscopy (AFM) were employed to study the micro- and nano-structure of V-amylose hosting LC fatty acids with different degrees of unsaturation. Additionally, enzymatic digestion tests were conducted to assess the release of the fatty acids from the complexes. X-ray diffraction confirmed the formation of V-amylose while light scattering measurements, and the AFM images revealed that increased fatty acid unsaturation lead to the formation of bigger and more disperse particle populations. Enzymatic digestion tests with mammalian pancreatin illustrated the control over guest release which is induced by enzymatic hydrolysis. In conclusion, this study demonstrates that guest chemistry affects some of the functional properties of V-amylose for the delivery of LC fatty acids.     

Short communication: Heritability of milk fatty acid composition and stearoyl-CoA desaturase indices in dairy cows

Activity of stearoyl-CoA desaturase (SCD) in the mammary gland is important for determining the relative proportions of saturated and monounsaturated fatty acids in milk and the concentration of the conjugated linoleic acid isomer rumenic acid (RA; cis-9,trans-11 18:2). Previous studies identified a large degree of between-cow variation in SCD activity, which was consistent across diets and suggests a genetic influence. The objectives of this study were to quantify genetic and phenotypic variations in fatty acid concentrations and SCD indices in milk fat and to estimate their heritabilities in a population of United Kingdom dairy cows. Milk samples were collected from 2,408 daughters of 597 Holstein-Friesian sires on 325 commercial farms for determination of fatty acid profiles. Indices of SCD activity were calculated by expressing each SCD product (cis-9 14:1, cis-9 16:1, cis-9 18:1, and RA) as a proportion of the precursor plus product [e.g., SCDI₁₄ = cis-9 14:1/(14:0 + cis-9 14:1)]. For individual fatty acids, phenotypic variance was considerably greater than additive genetic variance, resulting in small and nonsignificant heritability estimates (± standard error) for all except 6:0 (h² = 0.27 ± 0.10), 8:0 (h² = 0.27 ± 0.09), 12:0 (h² = 0.13 ± 0.07), cis-9 14:1 (h² = 0.28 ± 0.10), and cis-9 18:1 (h² = 0.12 ± 0.07). Heritability estimates of desaturase indices were significant for SCDI₁₄ (h² = 0.38 ± 0.11), SCDI₁₈ (h² = 0.19 ± 0.09), and SCDI_RA (h² = 0.21 ± 0.09), but not for SCDI₁₆ (h² = 0.05 ± 0.06). This study provides evidence that additive effects are responsible for a significant proportion of the phenotypic variation in SCD activity in dairy cows. It is concluded that because heritability of desaturase indices is moderate and significant in many cases, these indices could be investigated further for use in future breeding programs to increase concentrations of monounsaturated fatty acids and RA while decreasing concentrations of saturated fatty acids in milk fat.     

Short communication: estimates of genetic variation of milk fatty acids in US Holstein cows

Interest in changing the milk fatty acid profile is growing. However, little is known about the genetic variability of milk fatty acids in the US Holstein population. Therefore, genetic parameters for milk fatty acids were estimated using a single-trait, mixed, linear animal model on 592 individual milk samples from 233 daughters of 53 sires in a cow herd genetically representative of the US Holstein population. Heritability (h²) and repeatability (r) estimates ± standard errors for yields of individual fatty acids ranged from 0.00 ± 0.08 (C4:0) to 0.43 ± 0.13 (C12:0) for heritabilities and from 0.21 ± 0.05 (C18:1) to 0.43 ± 0.05 (C12:0) for repeatabilities. Saturated (h² = 0.23 ± 0.12; r = 0.36 ± 0.05) and de novo synthesized fatty acids (C6:0 to C14:0; h² = 0.30 ± 0.13; r = 0.40 ± 0.05) had numerically higher estimates than did monounsaturated (h² = 0.09 ± 0.09; r = 0.22 ± 0.05) and polyunsaturated fatty acids (h² = 0.08 ± 0.09; r = 0.27 ± 0.05). For relative proportions of individual fatty acids, the greatest heritability and repeatability estimates were obtained for C8:0 (h² = 0.18 ± 0.12; r = 0.36 ± 0.05), C10:0 (h² = 0.22 ± 0.13; r = 0.46 ± 0.05), C12:0 (h² = 0.18 ± 0.12; r = 0.46 ± 0.05), C16:0 (h² = 0.09 ± 0.12; r = 0.48 ± 0.05), C16:1 (h² = 0.49 ± 0.13; r = 0.49 ± 0.05), and C18:0 (h² = 0.24 ± 0.11; r = 0.39 ± 0.05). Our results suggest the existence of genetic variability of milk fatty acids, in particular of medium-and long-chain fatty acids (C8:0 to C18:0), which could be used to improve the nutritional and textural properties of milk fat by selective breeding.     

Identification of antioxidant components and fatty acid profiles of the leaves and fruits from Averrhoa carambola

The antioxidant compounds of leaves and fruits from Averrhoa carambola were extracted with acetone:water (7:3, v/v) and distilled water. The acetone:water extracts (AWE) were separated into five different subfractions, three obtained by liquid–liquid extraction using n-hexane (HF), ethyl acetate (EAF) and water (WF), and the other two obtained by further fractionation of the WF on a Sephadex LH-20 column (WS1 and WS2). The antioxidant activities, total phenolics and total flavonoids of these extracts and fractions were investigated. Results showed that all the extracts and fractions possessed potent antioxidant activities, as measured by DPPH, ABTS and FRAP methods, with the strongest for WS2 fractions accompanied with the highest total phenolics and total flavonoids. Reversed-phase HPLC, normal-phase HPLC, MALDI-TOF-MS and ¹³C NMR analyses identified that the main antioxidant components present in WS2 fractions were essentially of procyanidin-type proanthocyanidins, consisting mainly of epicatechin units linked by B-type interflavan bonds. In addition, GC–MS analysis revealed that the most abundant fatty acids were α-linolenic acid (62.04 ± 1.65%) for leaves and oleic acid (55.44 ± 1.37%) for fruits. The amount of total unsaturated fatty acids in leaves and fruits comprised more than 77% of total fatty acid.     

Nondestructive estimation of fatty acid composition in soybean [Glycine max (L.) Merrill] seeds using Near-Infrared Transmittance Spectroscopy

The potential of Near-Infrared Transmittance (NIT) Spectroscopy for estimation of fatty acid composition in soybean seed samples was studied. Total 612 whole seed samples with wide range of variability for major fatty acids were used to develop calibration equations by applying SNV de-trend and first derivative mathematical treatment in the range of 850–1048 nm. Useful chemometric models for most important fatty acids present in soybean seed oil were developed using Modified Partial Least Squares (MPLS) regression method. In external validation oleic (r² = 0.89, SEP = 1.61), linoleic (r² = 0.86, SEP = 1.50) and palmitic (r² = 0.89, SEP = 0.17) acids were predicted with good accuracy, while the predictions for linolenic acid (r² = 0.78, SEP = 0.36) and stearic acid (r² = 0.63, SEP = 0.11) had relatively poor accuracy. The whole-seed NIT spectroscopy equations for fatty acid estimation would be useful for improving efficiency of breeding programs aimed at altering fatty acid composition in soybean.     

Growth Inhibition of Mastitis Pathogens by Long-Chain Fatty Acids

Staphylococcus aureus, Staphylococcus hyicus, Streptococcus agalactiae, and Corynebacterium bovis were tested for sensitivity to long-chain fatty acids predominant in teat canal keratin. Anti-bacterial activity of free fatty acids on each bacterial species was measured after 12 and 24 h in chemically defined media. Polyene C_18:2 and C_18:3 acids were bactericidal to each species at ⩽10⁵ ng/ml⁻¹. The most bacteriostatic saturated fatty acids were C₁₂ and C₁₄. Streptococcus agalactiae growth was inhibited more by fatty acids after 24 h than after 12 h. No incubation time effect on growth responses of other species was determined. Polyoxyethylenesorbitan monooleate had a neutralizing effect on the bactericidal activity of polyene acids on C. bovis. Corynebacterium bovis were unable to grow in synthetic media containing individual free fatty acids as the sole source of preformed fatty acids. A relationship between bacterial species commonly isolated from bovine teat canals with resistance to fatty acids predominant in keratin was not evident.     

Retarding effects of organic acids,hydrocolloids and microwave treatment on the discoloration of green tea fresh noodles

Superfine green tea powder (SGTP) was premixed with organic acids (ascorbic acid, citric acid) and hydrocolloids (sodium alginate, curdlan), and then mixed with microwave-treated wheat flour to produce green tea fresh noodles (GTFN). Darken-retardant effects of organic acids, hydrocolloids and microwave treatments on GTFN were evaluated, as well as pH, polyphenol oxidase activity, sensory and microstructure characteristics. The results revealed that organic acids exhibited a suppressive effect on discoloration, among which citric acid (CA) displayed more efficient influence with lower pH. After adding hydrocolloids and microwave treatments, retardant effects exhibited more significant (P < 0.05). Specifically, employing citric acid 0.6 g/100 g, sodium alginate 0.2 g/100 g, and 800 W microwave (MW) 50 s would contribute to lower darkening index ΔE^* (24 h, 25 °C) at 3.88 ± 0.314, 4.94 ± 0.297, 2.78 ± 0.212, respectively. Furthermore, the combined effect of the above process restrained discoloring rate considerably (ΔE^* = 1.92 ± 0.101), also provided pleasant sensory characteristics. The confocal scanning laser microscopy (CSLM) images demonstrated the microstructure of the noodle was strengthened compared with blank GTFN, and sodium alginate could serve as a binding agent to parcel SGTP and starch granules.     

Triglycerides constituted of short and medium chain fatty acids and dicarboxylic acids in Momordica charantia, as well as capric acid,inhibit PGE2 production in RAW264.7 macrophages

This study was aimed to identify compounds in bitter gourd (Momordica charantia) ethyl acetate extract (EAE), which inhibit lipopolysaccharide-induced prostaglandin E₂ (PGE₂) production in RAW264.7 cells. Bitter gourd EAE was partitioned between n-hexane and methanol/H₂O (90/10). The hexane fraction was further separated by repeated silica gel chromatographies, and a reverse phase (RP) C18 chromatography. Fraction RP-10 showed the highest inhibition effect on PGE₂ production (Max inhibition = 96%, IC₅₀ = 2.3 μg/ml) and was identified to be triglycerides constituted of short and medium chain fatty acids by ¹H NMR, IR and H–HCOSY, and dicarboxylic acids by GC/MS. Fatty acids with 3–20 carbons were tested for the inhibitory activity, and capric acid exhibited the highest effect (Max inhibition = 99%, IC₅₀ = 6.5 μM). In conclusion, triglycerides composed of short and medium chain fatty acids and dicarboxylic acids in bitter gourd inhibit PGE₂ production, and capric acid is the most potent inhibitor among the fatty acids.     

Glycine max (L.) Merr., Vigna radiata L. and Medicago sativa L. sprouts: A natural source of bioactive compounds

The consumption of sprouts, common in Asia, has been growing in western countries, once they are a natural healthy food and considered as a valuable dietary supplement. Comparing with their mature counterparts, sprouts are usually richer in health-promoting phytochemicals. So, the nutritional composition and the biological potential of widely consumed sprouts of three species – Glycine max (L.) Merr., Vigna radiata L. and Medicago sativa L. – were compared for the first time. Phenolic compounds and phytosterols were analyzed by HPLC–DAD and organic acids by HPLC–UV. The volatile profile was determined by HS-SPME/GC–IT/MS. Fourteen phenolic compounds (including four isoflavones), three sterols one triterpene, sixteen fatty acids, seven organic acids and thirty volatile compounds were determined. The antioxidant activity was assessed against DPPH^?, superoxide and nitric oxide radicals. G. max sprouts were the most active against DPPH^? (IC₅₀ = 1.337 mg/mL), while those of M. sativa were the most effective against superoxide and nitric oxide radicals (IC₅₀ = 67 μg/mL and IC₅₀ = 426 μg/mL, respectively). Data provide evidence of great similarities between G. max and M. sativa sprouts, both being rich in phenolic compounds, fatty acids and volatiles, and exhibiting better antioxidant activity. On the other hand, V. radiata showed higher amounts of sterols, triterpenes and organic acids. In this study it was found that the sprouts are a good source of bioactive compounds in our diet with health-promoting antioxidants.     

Preparation of eicosapentaenoic acid concentrates from sardine oil by Bacillus circulans lipase

An extracellular lipase derived from Bacillus circulans, isolated from marine macroalga, Turbinaraia conoides, was used to prepare n-3 polyunsaturated fatty acid (PUFA) concentrates from sardine oil triglycerides. The enzyme was purified 132-fold with specific activity of 386 LU/mg. The purified lipase was able to enrich sardine oil with 37.7 ± 1.98% 20:5n-3 and 5.11 ± 0.14% 18:3n-3 in the triglyceride fraction after 3 h of hydrolysis. Lower hydrophobic constants of n-3 fatty acids (18:3n-3_logP = 5.65; 20:5n-3_logP = 5.85, respectively) than n-6 (20:4n-6_logP = 6.16) resulted in higher hydrolytic resistance of the former toward lipase, leading to their enrichment in the triglyceride fraction. Lipase-catalysed hydrolysis of sardine oil for 3 h, followed by urea complexation, provided free fatty acids containing 51.3 ± 4.65% 20:5n-3. The purified methyl ester of 20:5n-3 (68.29 ± 2.15%) from the urea concentrate was attained by chromatography on argentated neutral alumina.     

Optimization of gelatine gel preparation from New Zealand hoki (Macruronus novaezelandiae) skins and the effect of transglutaminase enzyme on the gel properties

Response Surface Methodology (RSM) was adopted to optimize the preparation of NZ hoki (Macruronus novaezelandiae) gelatine gels treated with Transglutaminase (TGase) and some of the rheological properties were characterized. The optimum concentration of the enzyme [X₁] was 3.33 mg/g, incubation time, [X₂] was 30 min, and incubation temperature, [X₃] was 37 ^°C. The calculated gel strength achieved by RSM was in very good agreement with the experimental value. The addition of TGase to hoki gelatine at the optimum concentration of increased the gel strength from 197 ± 5 g to 278.2 ± 0.19 g and the melting point from 21.4 ± 0.8 ^°C to 25.9 ± 0.1 ^°C. The increase in the G' values with the addition of TGase indicated the formation of firmer gels and the changes in G' and G" values with increase in temperature showed increase in melting point.     

C NMR and electrospray ionization mass spectrometric study of sucrose aqueous solutions at high pH: NMR measurement of sucrose dissociation constant

The ¹³C NMR technique is used for the measurement of the first dissociation constant of sucrose (HL) in highly alkaline solutions. In 1.0 M NaCl/NaOH medium and for 25 °C, the concentration dissociation constant (pK₁) was 13.1 ± 0.3; and, for 60 °C, pK₁ = 12.30 ± 0.05. The β-d-fructofuranosyl ring was found to be responsible for dissociation. The NMR data reveal no clear evidence of the second dissociation step below pH 14, either at 25 °C or at 60 °C. In the solutions with 4–10 mol dm⁻³ NaOH content the ¹³C NMR technique indicated the chemical shift changes, treated as the second dissociation step of sucrose and a sodium complex formation. A very rough estimation, for variable ionic strength, gives the value: pK₂ ∼ 15.8 ± 0.8. The anionic species L⁻ and NaH₋₁L⁻ have been registered by electrospray ionization time-of-flight mass spectrometry (ESI-ToF MS) for 0.01 M sucrose solutions with initial pH 13.     

Fatty acids profile of selected Artemisia spp. plants: Health promotion

In the present study we report the fatty acids profile of thirteen species of Artemisia, a hardy herb or shrub, analyzed by gas chromatography connected to a mass detector (GC-MS) for their nutritional value and their potential exploitation as a new source of essential fatty acids. Total lipids content ranged from 3.31 ± 0.19 to 17.78 ± 0.27 mg/g (fresh weight). The three most abundant fatty acids were C16:0, C18:2ω6 and C18:3ω3. Unsaturated fatty acids predominated in all the Artemisia species are studied with the α-linolenic acid (ALA) and linoleic acid (LA), which are essential for normal human growth, health promotion, and disease prevention. The predominant ω3 PUFA acid in all Artemisia species analyzed, was linolenic acid, with Artemisia gmellini, Artemisia ludoviciana and Artemisia vulgaris, showing higher amounts of this fatty acid, all thirteen species, analyzed in this study, were also rich in oleic acid (ω9) and linoleic acid (ω6), accounted for 50-70% of total PUFA. The ratio of ω3 PUFA to ω6 PUFA was similar in all species, varying from 1.0 to 3.0. Identifying Artemisia species as newer sources of PUFAs and enriching or optimizing the ω3FAs in known plant sources offer us ways of increasing the availability of ω3FAs in the food supply.     

Interaction of milk α- and β-caseins with tea polyphenols

The interaction of α- and β-caseins with tea polyphenols (+)-catechin (C), (−)-epicatechin (EC), (−)-epigallocatechin (EGC) and (−)-epigallocatechin gallate (EGCG) was examined at a molecular level, using FTIR, UV–visible, CD and fluorescence spectroscopic methods as well as molecular modelling. The polyphenol binding mode, the binding constant and the effects of polyphenol complexation on casein stability and conformation were determined. Structural analysis showed that polyphenols bind casein via both hydrophilic and hydrophobic interactions with overall binding constants of K_C–α-cas = 1.8 (±0.8) × 10³ M⁻¹, K_EC–α-cas = 1.8 (±0.6) × 10³ M⁻¹, K_EGC–α-cas = 2.4 (±1.1) × 10³ M⁻¹ and K_{EGCG–α-cas} = 7.4 (±0.4) × 10³ M⁻¹, K_C–β-cas = 2.9 (±0.3) × 10³ M⁻¹, K_EC–β-cas = 2.5 (±0.6) × 10³ M⁻¹, K_EGC–β-cas = 3.5 (±0.7) × 10³ M⁻¹ and K_{EGCG–β-cas} = 1.59 (±0.2) × 10⁴ M⁻¹. The number of polyphenol bound per protein molecule (n) was 1.1 (C), 0.9 (EC), 1.1 (EGC), 1.5 (EGCG) for α-casien and 1.0 (C), 1.0 (EC), 1.1 (EGC) and 1.5 (EGCG) for β-casein. Structural modelling showed the participation of several amino acid residues in polyphenol–protein complexation with extended H-bonding network. Casein conformation was altered by polyphenol with a major reduction of α-helix and β-sheet and increase of random coil and turn structure suggesting further protein unfolding. These data can be used to explain the mechanism by which the antioxidant activity of tea compounds is affected by the addition of milk.     

Study the effect of sun, oven and microwave drying on quality of onion slices

Sun, oven (50 and 70 °C) and microwave oven (210 and 700 W) drying of onion slices were carried out to monitor the drying kinetics and quality degradation of the product. Page, “Modified Page” and “Midilli and Küçük” models exhibited high coefficient of determination (R²) values, ranging between 0.994 and 0.999. The calculated effective diffusivity (D_eff) values (m²/s) of onion slices for the sun, oven 50 °C and oven 70 °C, microwave 210 W and microwave 700 W drying process were 8.339 × 10⁻¹⁰, 7.468 × 10⁻¹⁰, 1.554 × 10⁻⁹, 4.009 × 10⁻⁸ and 4.869 × 10⁻⁸, respectively. Fresh and dried onion slices had high amounts of K (696.82-16357.55 mg/kg), Ca (69.64-340.03 mg/kg), Na (37.72-1895.43 mg/kg), Mg (3.31-964.77 mg/kg) and P (46.47-3384.07 mg/kg) minerals. The highest mineral values were determined in oven dried samples. Sun (L^∗ 58.00 ± 4.83, a^∗ 0.27 ± 0.10, b^∗ 14.36 ± 2.40) and microwave oven drying (210 W) (L^∗ 54.78 ± 7.54, a^∗ −0.71 ± 0.09, b^∗ 13.17 ± 1.05) revealed better colour values in the dried products. The phenolic contents of microwave oven dried samples (1664.39 ± 134.12 and 1623.59 ± 140.02 for 210 W and 700 W, respectively) were higher than those of the other dried onion slices.     

Potentials to differentiate milk composition by different feeding strategies

To investigate the effect of the dietary intake of the cow on milk composition, bulk-tank milk was collected on 5 occasions from conventional (n = 15) and organic (n = 10) farms in Denmark and on 4 occasions from low-input nonorganic farms in the United Kingdom, along with management and production parameters. Production of milk based on feeding a high intake of cereals, pasture, and grass silage resulted in milk with a high concentration of α-linolenic acid (9.4 ± 0.2 mg/kg of fatty acids), polyunsaturated fatty acids (3.66 ± 0.07 mg/kg of fatty acids), and natural stereoisomer of α-tocopherol (RRR-α-tocopherol, 18.6 ± 0.5 mg/kg of milk fat). A milk production system using a high proportion of maize silage, by-products, and commercial concentrate mix was associated with milk with high concentrations of linoleic acid (LA; 19.7 ± 0.4 g/kg of fatty acids), monounsaturated fatty acids (27.5 ± 0.3 mg/kg of fatty acids), and a high ratio between LA and α-linolenic acid (4.7 ± 0.2). Comparing these 2 production systems with a very extensive nonorganic milk production system relying on pasture as almost the sole feed (95 ± 4% dry matter intake), it was found that the concentrations of conjugated LA (cis-9,trans-11; 17.5 ± 0.7 g/kg of fatty acids), trans-11-vaccenic acid (37 ± 2 g/kg of fatty acids), and monounsaturated fatty acids (30.4 ± 0.6 g/kg of fatty acids) were higher in the extensively produced milk together with the concentration of antioxidants; total α-tocopherol (32.0 ± 0.8 mg/kg of milk fat), RRR-α-tocopherol (30.2 ± 0.8 mg/kg of milk fat), and β-carotene (9.3 ± 0.5 mg/kg of milk fat) compared with the organic and conventional milk. Moreover, the concentration of LA (9.2 ± 0.7 g/kg of fatty acids) in milk from the extensive milk production system was found to approach the recommended unity ratio between n-6 and n-3, although extensive milk production also resulted in a lower daily milk yield.     

Microbiological quality of fresh lettuce from organic and conventional production

Previously there was no available information on the levels of indicator bacteria and the prevalence of pathogens in fresh lettuce grown in organic and conventional farms in Spain. A total of 72 lettuce samples (18 farms for 4 repetitions each) for each type of the agriculture were examined in order to assess the bacteriological quality of the lettuces, in particular the prevalence of selected pathogens. The lettuce samples were analyzed for the presence of aerobic mesophilic, psychrotrophic microorganisms, yeasts and moulds, Enterobacteriaceae, mesophilic lactic acid bacteria, Pseudomonas spp. and presumptive Escherichia coli, Salmonella spp. and Listeria monocytogenes. The mean aerobic mesophilic counts (AM) were 6.35 ± 0.69 log₁₀ cfu g⁻¹ and 5.67 ± 0.80 log₁₀ cfu g⁻¹ from organic and conventional lettuce, respectively. The mean counts of psychrotrophic microorganisms were 5.82 ± 1.01 log₁₀ cfu g⁻¹ and 5.41 ± 0.92 log₁₀ cfu g⁻¹ from organic and conventional lettuce, respectively. Yeasts and moulds (YM) mean counts were 4.74 ± 0.83 log₁₀ cfu g⁻¹ and 4.21 ± 0.96 log₁₀ cfu g⁻¹ from organic and conventional lettuce, respectively. Lactic acid bacteria (LAB) were present in low numbers and the mean counts were 2.41 ± 1.10 log₁₀ cfu g⁻¹ and 1.99 ± 0.91 log₁₀ cfu g⁻¹ from organic and conventional lettuce, respectively. Pseudomonas spp. mean counts were 5.49 ± 1.37 log₁₀ cfu g⁻¹ and 4.98 ± 1.26 log₁₀ cfu g⁻¹ in organic and conventional lettuce, respectively. The mean counts for Enterobacteriaceae were 5.16 ± 1.01 log₁₀ cfu g⁻¹ and 3.80 ± 1.53 log₁₀ cfu g⁻¹ in organic and conventional lettuce, respectively. E. coli was detected in 22.2% (16 samples) of organic lettuce and in 12.5% (9 samples) of conventional lettuce. None of the lettuce samples was positive for E. coli O157:H7, L. monocytogenes and Salmonella spp. From the samples analyzed by principal component analysis (PCA) a pattern with two different groups (conventional and organic) can be observed, being the highest difference between both kinds of samples the Enterobacteriaceae count.     

In vitro fermentation of polysaccharide from the seeds of Plantago asiatica L. by human fecal microbiota

In vitro fermentation of the polysaccharide (PLCP, Mw = 1.90 × 10⁶ Da) from seeds of Plantago asiatica L. and the contribution of its carbohydrates to the fermentation was investigated in this study. The polysaccharide was characterized by high contents of xylose, arabinose and glucuronic acid, and it was subjected to human fecal cultures to be fermented in vitro for 24 h. During fermentation, pH in fecal cultures decreased from 6.1 to 5.1 and the levels of total short-chain fatty acid (SCFA), acetic, propionic and n-butyric acids all significantly increased. Xylanase, arabinofuranosidase, xylosidase and glucuronidase activities were also improved. After 24 h incubation, 47.2 ± 1.6% of total carbohydrate in polysaccharide, including 42.9 ± 1.5% of arabinose, 53.2 ± 1.6% of xylose and 76.4 ± 1.2% of glucuronic acid, were consumed. In addition, relationship between carbohydrate consumption of the polysaccharide and SCFA production was also evaluated. It was found that the increase of acetic and n-butyric acid productions mainly resulted from the fermentation of glucuronic acid and xylose in polysaccharide, while the increase of propionic acid production was primarily due to the fermentation of arabinose and xylose. These results showed that the polysaccharide was physiologically active for human large bowel, and its carbohydrate composition determined its SCFA production.