Use of UFLC-PDA for the Analysis of Organic Acids in Thirty-Five Species of Food and Medicinal Plants

Analysis of organic acids has become increasingly important due to their role in the physiological activity of plants, and many separation methods have been developed for the simultaneous determination of these compounds in plant samples. Herein, ultra fast liquid chromatography and photodiode array detection (UFLC-PDA) was applied to the analysis of organic acids in young shoots, leaves, aerial parts and flowering shoots, as well as in flowers and fruits, of 35 plant species, according to their traditional use. The studied plants were divided in three groups: traditionally cultivated food plants, wild edible plants and wild medicinal plants. Most of the species were characterized for the first time. Among all the analysed species, Rumex acetosella leaves and aerial parts revealed the highest content of total organic acids. Overall, the organic acids found in the studied plant species make them suitable to be used as food additives such as antioxidants (e.g. ascorbic acid) or acidulants (e.g. citric and malic acids).     

Composition of Sugars,Organic Acids,and Total Phenolics in 25 Wild or Cultivated Berry Species

Abstract: Sugars, organic acids, and total phenolic content in fruit of 25 wild and cultivated berry species were identified and quantified with high‐performance liquid chromatograph. The composition of sugars, organic acids, and total phenolic compounds in various species of Vaccinium, Rubus, Ribes, and Fragaria genus was evaluated. Additonally, total phenolics of less known berry species of the Morus, Amelanchier, Sorbus, Sambucus, Rosa, Lycium, Actinidia, and Aronia genus were determined in wild growing as well as in cultivated fruits. Significant differences in the concentration of sugars and organic acids were detected among the berry species. Glucose and fructose were the most abundant sugars in berry fruits and the major organic acids were malic and citric acid. However, in kiwi fruit, sucrose represented as much as 71.9% of total sugars. Sorbitol has been detected and quantified in chokeberry, rowanberry, and eastern shadbush fruit. The highest content of total analyzed sugars was determined in rowanberry fruit, followed by dog rose, eastern shadbush, hardy kiwifruit, American cranberry, chokeberry, and jostaberry fruit. Rowanberry stands out as the fruit with the highest content of total analyzed organic acids, followed by jostaberry, lingonberry, red gooseberry, hardy kiwifruit, and black currant. The berries of white gooseberry, black currant, red currant, and white currant had the lowest sugar/organic acid ratio and were thus perceptively the sourest species analyzed. On the other hand, the species with highest sugar/organic acid ratio were goji berry, eastern shadbush, black mulberry, and wild grown blackberry. The highest amounts of total phenols were quantified in chokeberry fruit. Wild strawberry, raspberry, and blackberry had 2‐ to 5‐fold more total phenolics compared to cultivated plants. Practical Application: The fruit of analyzed berry species contained different levels of sugars, organic acids, and total phenolics. Moreover, it has been demonstrated that wild grown species generally contain more phenolics than cultivated ones. This information is interesting for nutritionists as well as berry growers and breeders who can promote the cultivation of species and new cultivars with higher phenolic content.     

Simultaneous quantification of caffeoyl esters and flavonoids in wild and cultivated cardoon leaves

Cynara cardunculus is a diploid (2n = 2x = 34) species, native to the Mediterranean basin, which belongs to the family of Asteraceae. It includes globe artichoke, cultivated cardoon, as well as their progenitor wild cardoon. The species is a source of biophenols and its leaf extracts have been widely used in herbal medicine as hepatoprotectors and choleretics since ancient times. Globe artichoke leaves have been found to be rich in compounds originating from the metabolism of phenylpropanoids however, to our knowledge, the leaf polyphenolic composition of the two other forms within the species, cultivated and wild cardoon, have not yet been properly investigated. Two main classes of polyphenols have been detected by HPLC/DAD and HPLC/MS analyses: caffeoyl esters and flavonoids. The compounds which are the result of esterification of caffeoylquinic acid moiety with succinic acid, previously detected in other members of the Asteraceae family, were detected in cardoon leaves for the first time.     

Oil Content and Fatty Acids Composition in Brassica Species

Seeds of 20 accessions of six Brassica species including cultivated and five wild relatives were analyzed for oil and fatty acid composition. The results showed that oil content varied from 21 (B. nigra) to 46% (B. napus). Among wild species, B. rapa and B. oleracea had highest oil content (31 and 28%, respectively). The main fatty acids of oleic, linoleic, linolenic, erucic, palmitic, and stearic acids accounted for 89–94% of the total fatty acids in all species. Cultivated species of B. napus had highest oleic acid (61%) and lowest erucic acid (1%) content compared to other studied species. Brassica rapa and B. oleracea had the highest content of erucic acid (41 and 46%, respectively). The highest content of linolenic (20%) and linoleic (19%) acid was observed for B. juncea. The results showed that there was high genetic variation among the studied species for oil content and fatty acids composition. This indicates that seed oil of these species is possibly suitable for both human consumption and industrial purposes.     

Polyphenolic composition and content in the ripe berries of wild Vitis species

To explore wild genetic resources for improving fruit and processing quality of cultivated grape cultivars, the polyphenolic composition and content in the ripe berries of 147 grape accessions from 16 Vitis species for two consecutive years were characterised. A total of 48 polyphenolic compounds, including 28 anthocyanins, 6 flavonols, 6 flavanols, 6 hydroxycinnamic derivatives, and 2 hydroxybenzoic acids, were identified via HPLC-MS and quantified by HPLC-DAD. These wild grape species had unique presence of abundant di-glucoside derivatives of anthocyanins. In addition, anthocyanins in most wild species were predominantly nonacylated. The mean contents for anthocyanins, flavanols, flavonols, hydroxycinnamic derivatives, and hydroxybenzoic acids were 9.610, 0.769, 0.093, 0.441 and 0.027 mg g⁻¹ FW, respectively. They were about 2 to 10 folds higher than their respective counterparts in the most widely cultivated grape species Vitis vinifera. As expected, most of these groups of compounds were correlated negatively with berry weight, but positively with the content of total soluble solids.     

Variation in seed protein content in the annual wild Cicer species

In a search for higher seed protein content than in cultivated chickpea (Cicer arietinum L), the authors evaluated a collection of 228 accessions from the International Center for Agricultural Research in the Dry Areas (ICARDA), representing the eight annual wild species of the genus Cicer, along with 20 cultivated chickpea check lines. Variation in seed protein content ranged from 168 g kg⁻¹ in C cuneatum Hochst ex Rich to 268 g kg⁻¹ in C pinnatifidum Jaub & Spach, with an average seed protein content of 207 g kg⁻¹ over the eight wild species. C yamashitae Kitamura had the highest mean (217 g kg⁻¹), while C echinospermum PH Davis had the lowest (192 g kg⁻¹). The mean protein content of the cultivated checks was 188 g kg⁻¹. Significant variation was present in C judaicum Boiss, C pinnatifidum and C reticulatum Ladiz. C pinnatifidum had the highest number of accessions with high protein content. Overall, protein content showed negative association with harvest index, but little association with leaf area, days to maturity and canopy width. Although the variation for seed protein content of the collection showed accessions with higher protein content than with the cultivated checks, it falls within the range reported for cultivated chickpea. It is expected that agronomically superior selections from interspecific hybrids involving C arietinum and its annual relatives should not be inferior to the cultigen in protein content. Moreover, as usually occurs in distant hybridisation, unexpected epistatic effects could produce positive transgressive segregants, as has already been reported in Cicer. © 1998 Society of Chemical Industry     

Chemical composition and in vitro digestibility of some Spanish browse plant species

Samples of leaves, flowers and fruits of four browse species were collected from uplands in the province of León (northwestern Spain) at different sampling times (from spring till autumn). The browse species were Erica australis (Spanish heath), Cistus laurifolius (laurel‐leaved rock rose), Quercus pyrenaica (hoary oak) and Rosa canina (wild dog rose). A large variability in chemical composition and in vitro digestibility and gas production kinetics was observed among species and among sampling times within each species. The ranking order of the browse plants according to their in vitro digestibility and gas production kinetics was R canina > Q pyrenaica > C laurifolius > E australis. In the leaves of R canina and Q pyrenaica, crude protein content, digestibility and parameters of gas production tended to decrease throughout the growing season (P < 0.05 in most cases), whereas cell wall contents followed the opposite trend. Seasonal variations were less pronounced in the other browse species. There were significant (P < 0.05) correlations between in vitro digestibility and chemical composition of the browse plants, positive with the crude protein content and negative with the fibre fractions. Copyright © 2004 Society of Chemical Industry     

Technology Allowing Baby Spinach Leaves to Acquire Freezing Tolerance

This study focuses on improving the freezing tolerance of spinach leaves when applying vacuum impregnation (VI) and pulsed electric fields (PEF) as pre-treatments. Changing the cultivation conditions of the spinach plants was tested for improved freezing tolerance of the harvested leaves. Spinach plants were initially cultivated at 20 °C for 5 weeks and harvested at the beginning of week 6 before the cultivation conditions were changed to 5 °C. After exposing the plants to cold stress, leaves were harvested on day 1, 8, and 20. The leaves from different cultivation temperatures were treated with VI with 30% (w/v) trehalose solution and PEF prior freezing at ??22 °C in a blast freezer. After freezing and thawing, the viability of harvested spinach leaves was 50% for the plants cultivated at 20 °C for 5 weeks. When the cultivation conditions were changed, the viability of the harvested leaves increased to 82% on day 1, 89% on day 8, and without a significant further increase on day 20. During cold cultivation, sucrose accumulated in the leaves, which might have contributed to the increased survival. The influence of impregnating the accumulated sugars instead of changing the cultivation conditions on the survival of the leaves was tested. The viability of the leaves was 75%, which was higher than the survival of the control (50%). The results indicate that it is possible to increase leaf survival after freezing and thawing by applying VI and PEF in combination with either by changing the cultivation conditions of spinach plants or by externally impregnating additional sugars to the harvested leaves.     

Effect of natural biostimulants on yield and nutritional quality: an example of sweet yellow pepper (Capsicum annuum L.) plants

BACKGROUND: Modifications in growing techniques can affect the yield and nutritional quality of various cultivated plant species. Owing to its high nutritional value, pepper (Capsicum annuum L.) was used in this study as a model plant to investigate the effect of natural biostimulants on yield and fruit quality parameters under conditions of reduced fertilisation. RESULTS: A positive influence of biostimulant treatment on yield parameters was observed. The overall increase in the pigment content of leaves after biostimulant application agreed well with the higher total and commercial yields of treated pepper cultivars compared with their controls. The results showed that natural biostimulants had a positive effect on the vitamin C and total phenolic contents in pepper fruits during the hot summer season. The 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) and 2,2′‐azinobis(3‐ethylbenzothiazoline‐6‐sulfonate) (ABTS) antioxidant activities were also significantly higher (P < 0.05) in treated plants and correlated strongly with all measured quality parameters except total phenolic content. CONCLUSION: Generally, biostimulants improved the antioxidant activity, vitamin C and phenolic contents in fruits as well as the pigment content in leaves of treated compared with non‐treated pepper plants grown hydroponically. Thus the application of biostimulants could be considered as a good production strategy for obtaining high yields of nutritionally valuable vegetables with lower impact on the environment. Copyright © 2011 Society of Chemical Industry     

CHEMICAL AND ANTIOXIDANT PROPERTIES OF CULTIVATED AND WILD FRAGARIA AND RUBUS BERRIES

ABSTRACT

Content of individual sugars, organic acids, total phenolics (TPH), some important phenolic compounds (kaempferol, myricetin, quercetin, and ellagic acid) and total antioxidant capacity (TAC) in the fruits of strawberry, raspberry and blackberry were studied. A comparison was made between cultivars and wild relatives of each species (Fragaria vesca L., Rubus idaeus L. and Rubus fruticosus L.). The main sugars found were fructose and glucose both in the fruits of wild species and the studied cultivars. Citric acid was determined to be the major organic acid in most of tested berries with the exception of blackberry cultivars, where malic acid was dominant. The content of individual phenolic compounds varied among the wild species, as well as among the studied cultivars. Ellagic acid content was higher in F. vesca, R. idaeus and R. fruticosus (122.5 µg/g FW, 12.71 µg/g FW and 61.7 µg/g FW, respectively) than that obtained in analyzed cultivars. Overall, TPH expressed higher values in the wild strawberry and blackberry species in comparison to the studied cultivars, and consequently, the highest levels of TAC were recorded in F. vesca (5.78 mg asc/g FW), followed by R. fruticosus (4.95 mg asc/g FW).

PRACTICAL APPLICATIONS

In recent years, both wild and cultivated berries have become very attractive for consumers because of potentially beneficial phytochemicals contained in these fruits. Fruit nutritional quality can be described by standard quality parameters (sugars and organic acids), and the analysis of antioxidant capacity influenced by specific related compounds. The importance of flavonoids and other phenolics have been suggested to play a preventive role in the development of cancer and heart disease. A significant positive correlation observed in this study between total phenolics and total antioxidant capacity indicate the need for the use of wild species in the breeding programs of small fruits, especially strawberry and blackberry, in order to increase their nutritive value and the health benefits.     

Plant location and extraction procedure strongly alter the antimicrobial activity of murta extracts

Leaves and fruits of Murta (Ugni Molinae Turcz.) growing in three locations of Chile with diverse climatic conditions were extracted by using ethanol/water mixtures at different ratios and the antimicrobial activity was assessed. Extracts containing the highest polyphenolic content were from murta plants grown nearer to the mountain (58 mg GAE/g murta), subjected to extreme summer/winter-day/night temperature changes and rainy regime. Extracts from leaves collected in the valley and coast contained 46 and 40 mg GAE/g murta, respectively. A mixture of 50% ethanol/water was the most efficient in extracting polyphenols, showing pure solvents—both water and ethanol—a lower extraction capacity. No correlation between antioxidant capacity and polyphenolic content was found. Extracts from Murta leaves provoked a decrease in the growing of Pseudomonas aeruginosa, Klebsiella pneumoniae and Staphylococcus aureus, and showed no activity against the beneficial, probiotic bacteria. A significant correlation between polyphenol content and antimicrobial activity on harmful bacteria was found. Myricetin glucoside and quercetin glucoside/glucuronide/dirhamnoside presumably contributed to the antimicrobial activity of the extract. The higher antimicrobial activity of leaves extracts compared to the fruits could be attributed to flavan-3-ols and other flavonol glycosides. Quercetin glucuronide, myricetin xyloside and flavan-3-ols in polymeric form were tentatively identified for the first time in murta extracts. Both extracts showed an antimicrobial activity similar to some commercial antibiotics, suggesting their suitability to replace synthetic antimicrobials in food.     

Peanut improvement: production of fertile hybrids and backcross progeny between Arachis hypogaea and A. kretschmeri

There are only a few reports of successful crosses between cultivated peanut (Arachis hypogaea L., section Arachis) and wild species from sections other than section Arachis. Many of the wild Arachis species harbor important traits necessary for the improvement of peanut. For example, Arachis kretschmeri Krapov., W.C. Gregory & C.E. Simpson (section Procumbentes) can grow under water-logged conditions and has been identified as one of the few wild species of Arachis with resistance to late leaf spot (LLS) and peanut rosette disease. Peanut rosette, caused by a combination of viruses, is an economically important disease only in Africa, while LLS, caused by Cercosporidium personatum, is an important fungal disease in Asia and the Americas as well as Africa. Interspecific hybrids between A. hypogaea and A. kretschmeri were produced by applying growth regulators to pollinated pistils and hybrid plants were obtained by germinating embryos in vitro. A total of seven hybrids were produced and confirmed by Simple Sequence Repeat (SSR) analysis. All hybrids were fertile, although initially slow growing. F₁ hybrids were backcrossed to A. hypogaea and all plants in the F₁BC₁ generation were single-seeded with a prominent beak, characteristic of A. kretschmeri, but many of the F₁BC₂ pods were double-seeded resembling A. hypogaea. F₁BC₂ plants were moderately resistant to LLS. When a large number of seeds are obtained, the progeny will be screened for resistance to both LLS and rosette disease. Thus crosses with species outside the section Arachis may not only confer disease resistance but will also broaden the genetic base of cultivated peanut.     

Seed decontamination as an intervention step for eliminating Escherichia coli on salad vegetables and herbs

The efficacy of seed decontamination to enhance the safety of salad vegetables and herbs was evaluated. Seeds (celery, coriander, lettuce, spinach and watercress) were inoculated (at a level of 3–5 log cfu g⁻¹) with either Escherichia coli P36 or Listeria monocytogenes NCTC 7973 and decontaminated with ozone gas, acidified sodium chlorite (ASC) or quaternary ammonium salt preparation (QAS). None of the treatments applied were initially effective at inactivating E coli on lettuce or spinach seeds as the bacterium could be recovered on the subsequent seedlings. However, as the cultivation period progressed, E coli numbers on plants derived from decontaminated seeds declined to below the level of detection. Interestingly, E coli persisted on the surface of lettuce and spinach leaves from inoculated non‐treated seeds throughout the 42‐day cultivation period. E coli also persisted on coriander derived from inoculated non‐treated seeds although it was isolated sporadically on plants derived from QAS‐decontaminated seed. E coli numbers progressively declined on celery and watercress regardless of the seed decontamination treatment being applied. No L monocytogenes was recovered from any of the seedlings, possibly because of the growth‐suppressing effect of endogenous microflora. The results suggest that effective on‐farm controls, such as seed decontamination, should be considered in order to reduce the risk of pathogens associated with salad vegetables and herbs. Copyright © 2005 Society of Chemical Industry     

Conventional,organic and biodynamic farming: differences in polyphenol content and antioxidant activity of Batavia lettuce

BACKGROUND: Lactuca sativa L. ssp. acephala L., cv. Batavia red Mohican plants were cultivated under intensive conventional, organic and biodynamic farming and were analyzed for their polyphenol content and antiradical activity in order to demonstrate the influence of farming on yield, polyphenol content and antiradical activity. RESULTS: The yield of plants from conventional farming was the highest (2.89 kg m^?2), while polyphenol content, measured by spectrophotometry, of these plants was lower at P < 0.05 (1.36 mg g^?1) than the content of plants from organic and biodynamic farming (1.74 and 1.85 mg g^?1, respectively). The antiradical activity, measured by DPPH · assay, was positively correlated to flavonoid and hydroxycinnamic acid contents. CONCLUSION: Flavonoid, hydroxycinnamic acid and anthocyan patterns were not affected by the type of cultivation, while quantitative differences were demonstrated and some differences were found between conventional farming and organic or biodynamic farming. The yield of conventionally grown salads was the highest. Copyright © 2011 Society of Chemical Industry     

Effect of High-Pressure Homogenization on the Physical and Antioxidant Properties of Quercus resinosa Infusions Encapsulated by Spray-Drying

Abstract: Quercus resinosa leaves are used in northern Mexico as a refreshing beverage rich in polyphenolic compounds. These leaves show astringency and hence need taste masking for incorporating in a food product. They also interact with many other food components and are not very stable to food processing environments, thus it is important to protect them and a common way is by encapsulation. In the present study the use of encapsulation by spray-drying of Quercus resinosa leaves infusions was evaluated. Q. resinosa leaves were collected, air dried, and milled prior to infusion preparation. Lactose–sodium caseinate blends at 3 different proportions (11 : 4%, 9 : 6%, and 7 : 8%) were dispersed with a constant amount of lyophilized infusion (0.075%) and processed under high-pressure homogenization (0, 100, 200, 300 MPa). Total phenolic content, DPPH* kinetic analysis, deoxy-D-ribose oxidation inhibition, rheological evaluation, and particle size analysis were performed to evaluate the obtained capsules. High antioxidant activity was shown by capsules despite their very low concentration when inhibiting deoxy-D-ribose oxidation. Chain breaking rate was related to polyphenolic concentration in capsules. Using lactose-caseinate blends produces capsules of submicron to nanometer size that retain the good antioxidant capacities of original infusions.     

Antioxidants in spring leaves of Oxalis acetosella L.

Oxalis acetosella L. is a common, edible wild plant native to the northern hemisphere. The contents of selected antioxidants, and the antioxidant capacity of young and old spring leaves of O. acetosella, were evaluated. The present study reports foliar contents of ascorbic acid, tocopherols, carotenoids, chlorophyll, flavonoids, phenolic acids and total phenolics, and compares the nutritional value of O. acetosella with other cultivated and wild plants. The composition of foliar antioxidants was found to depend on leaf age. On the other hand, the antioxidant capacity of old leaves were in the same range as young leaves. A comparison between O. acetosela with lettuce analysed in our study, and with numerous cultivated and wild edible plants from other studies, showed that O. acetosella is very rich in β-carotene, ascorbic acid, tocopherols and xanthophylls, and that it is one of the best sources of flavonoids (flavonol glycosides and flavan-3-ols), especially rutin. Therefore, O. acetosella is a potentially important dietary source of antioxidants.     

Aluminium in wild mushrooms and cultivated Agaricus bisporus

 The aluminium content in wild mushrooms (n = 271, 19 species) and in cultivated Agaricus bisporus (n = 15) was determined by graphite furnace atomic absorption spectroscopy. With an aluminium content of 30 – 50 μ/g dry matter (DM) Boletus and Xerocomus species, the most well-known and most popular mushrooms, proved to be poor in aluminium. Several species of the genus Suillus, Macrolepiota rhacodes, Hypholoma capnoides as well as individual samples of Russula ochroleuca and Amanita rubescens contained high aluminium concentrations of about 100 μg/g DM and more. Cultivated Agaricus bisporus had the lowest aluminium content, i. e. 14 μg/g DM. The site, its geological origin as well as the mushroom species influenced the aluminium content in the fruitbodies: these factors require further investigation. Mushrooms do not contribute significantly to aluminium intake by humans. Received: 23 January 1997     

Ecological factors associated with pest status in Callosobruchus (Coleoptera: Bruchidae): high host specificity of non-pests to Cajaninae (Fabaceae)

Larval host plants of six Callosobruchus species (Coleoptera: Bruchidae), including pest species, were discovered by collecting the seeds of wild (or inedible) and cultivated edible legumes in the field in Taiwan, Thailand, continental China, Myanmar, Nepal and the Philippines. A close relationship between Asian Callosobruchus species and the leguminous subtribe Cajaninae was revealed: Rhynchosia species were commonly used by three Callosobruchus species, wild and cultivated species of Cajanus and two Dunbaria species, respectively, by single species. Two Taiwanese species were confirmed to be conspecific with continental species. We further reviewed host legumes of 11 species of Callosobruchus including nine species of pests. There were significant positive correlations between geographic distribution range and host range at all three taxonomic levels of hosts. Principal component analysis on geographic distribution range, host range, altitude of distribution, utilizations of cultivated hosts, of Cajaninae and of Phaseolinae showed that the first axis (PC1) described 52% of total variance, which was related significantly with the frequencies of utilization of cultivated legumes (0.93), and of Cajaninae (Cajanus, Dunbaria and Rhynchosia) (−0.85). PC1 was also positively correlated with the frequency of utilization of Phaseolinae (Vigna and Lablab) (0.68), geographical range (0.67) and with host range (0.67) before Bonferroni corrections. Contrary to the polyphagy of the widely distributed pest Callosobruchus, non-pest species exhibited fidelity to single specific genera of wild or inedible legumes, and pests with limited distribution are specific to leguminous subtribes. Non-pests are characterized by tight association with Cajaninae. We concluded that specialization to non-economic Cajaninae prevents a species of Callosobruchus from becoming a pest of cultivated legumes.     

Microbiological quality and safety of fresh cultivated and wild mushrooms commercialized in Spain

402 samples of 22 species of cultivated and wild fresh mushrooms sold in retail markets and supermarkets in Zaragoza (Spain) were studied to quantify their microbial load (mesophilic aerobic microorganisms, Pseudomonas genus, Enterobacteriaceae, lactic acid bacteria, total and thermotolerant coliform bacteria, Escherichia coli, yeasts and moulds) and to investigate the presence of E. coli O157:H7, Listeria monocytogenes, Salmonella spp., Staphylococcus aureus and Yersinia enterocolitica. The total microbial counts ranged from 4.4 to 9.4 log cfu/g, the genus Pseudomonas being the most prevalent with counts from 3.7 to 9.3 log cfu/g and Auricularia auricula-judae the species with the highest microbial load (9.4 log cfu/g). No significant differences (p > 0.05) were detected between mean counts of wild and cultivated species in all the microbial groups studied. The microbiological safety level of the cultivated mushrooms was excellent since no pathogens were isolated, and the microbial counts of indicator microorganisms were low, being detected in only half of the species. Salmonella spp, E. coli O157:H7 and S. aureus were not isolated from any sample, Y. enterocolitica was detected in only four samples of wild mushrooms whereas twenty-six (6.5%) were positive for L. monocytogenes, their occurrence being relatively high in Calocybe gambosa (40%), Hygrophorus limacinus (40%) and Tuber indicum (100%). These results suggest that a strategy to reduce bacterial populations, and to improve the microbiological safety of some species of fresh mushrooms, should be investigated.