Culture Dependent and Independent Analysis of Potential Probiotic Bacterial Genera and Species Present in the Phyllosphere of Raw Eaten Produce.

The plant phyllosphere is colonized by a complex ecosystem of microorganisms. Leaves of raw eaten vegetables and herbs are habitats for bacteria important not only to the host plant, but also to human health when ingested via meals. The aim of the current study was to determine the presence of putative probiotic bacteria in the phyllosphere of raw eaten produce. Quantification of bifidobacteria showed that leaves of Lepidium sativum L., Cichorium endivia L., and Thymus vulgaris L. harbor between 103 and 106 DNA copies per gram fresh weight. Total cultivable bacteria in the phyllosphere of those three plant species ranged from 105 to 108 CFU per gram fresh weight. Specific enrichment of probiotic lactic acid bacteria from C. endivia, T. vulgaris, Trigonella foenum-graecum L., Coriandrum sativum L., and Petroselinum crispum L. led to the isolation of 155 bacterial strains, which were identified as Pediococcus pentosaceus, Enterococcus faecium, and Bacillus species, based on their intact protein pattern. A comprehensive community analysis of the L. sativum leaves by PhyloChip hybridization revealed the presence of genera Bifidobacterium, Lactobacillus, and Streptococcus. Our results demonstrate that the phyllosphere of raw eaten produce has to be considered as a substantial source of probiotic bacteria and point to the development of vegetables and herbs with added probiotic value.

Mycotoxins in spices and herbs-An update.

Spices and herbs have been used since ancient times as flavor and aroma enhancers, colorants, preservatives, and traditional medicines. There are more than 30 spices and herbs of global economic and culinary importance. Among the spices, black pepper, capsicums, cumin, cinnamon, nutmeg, ginger, turmeric, saffron, coriander, cloves, dill, mint, thyme, sesame seed, mustard seed, and curry powder are the most popular spices worldwide. In addition to their culinary uses, a number of functional properties of aromatic herbs and spices are also well described in the scientific literature. However, spices and herbs cultivated mainly in tropic and subtropic areas can be exposed to contamination with toxigenic fungi and subsequently mycotoxins. This review provides an overview on the mycotoxin risk in widely consumed spices and aromatic herbs.

Traditional food uses of wild plants among the Gorani of South Kosovo.

A food ethnobotanical field study was conducted among the Gorani of South Kosovo, a small ethnic minority group that speaks a South-Slavic language and lives in the south of the country. We conducted forty-one semi-structured interviews in ten villages of the Kosovar Gora mountainous area and found that seventy-nine wild botanical and mycological taxa represent the complex mosaic of the food cultural heritage in this population. A large portion of the wild food plant reports refer to fermented wild fruit-based beverages and herbal teas, while the role of wild vegetables is restricted. A comparison of these data with those previously collected among the Gorani living in nearby villages within the territory of Albania, who were separated in 1925 from their relatives living in present-day Kosovo, shows that approximately one third of the wild food plant reports are the same. This finding demonstrates the complex nature of Kosovar Gorani ethnobotany, which could indicate the permanence of possible "original" Gorani wild plant uses (mainly including wild fruits-based beverages), as well as elements of cultural adaptation to Serbian and Bosniak ethnobotanies (mainly including a few herbal teas and mushrooms).

UFLC-ESI-MS/MS analysis of multiple mycotoxins in medicinal and edible Areca catechu.

A robust, sensitive and reliable ultra fast liquid chromatography combined with electrospray ionization tandem mass spectrometry (UFLC-ESI-MS/MS) was optimized and validated for simultaneous identification and quantification of eleven mycotoxins in medicinal and edible Areca catechu, based on one-step extraction without any further clean-up. Separation and quantification were performed in both positive and negative modes under multiple reaction monitoring (MRM) in a single run with zearalanone (ZAN) as internal standard. The chromatographic conditions and MS/MS parameters were carefully optimized. Matrix-matched calibration was recommended to reduce matrix effects and improve accuracy, showing good linearity within wide concentration ranges. Limits of quantification (LOQ) were lower than 50 µg kg(-1), while limits of detection (LOD) were in the range of 0.1-20 µg kg(-1). The accuracy of the developed method was validated for recoveries, ranging from 85% to 115% with relative standard deviation (RSD) ≤14.87% at low level, from 75% to 119% with RSD ≤ 14.43% at medium level and from 61% to 120% with RSD ≤ 13.18% at high level, respectively. Finally, the developed multi-mycotoxin method was applied for screening of these mycotoxins in 24 commercial samples. Only aflatoxin B2 and zearalenone were found in 2 samples. This is the first report on the application of UFLC-ESI(+/-)-MS/MS for multi-class mycotoxins in A. catechu. The developed method with many advantages of simple pretreatment, rapid determination and high sensitivity is a proposed candidate for large-scale detection and quantification of multiple mycotoxins in other complex matrixes.

Lipopolysaccharide IP-PA1 from Pantoea agglomerans prevents suppression of macrophage function in stress-induced diseases.

Chronic psychological stress impairs health and induces various diseases by causing an imbalance in the immune, neuropsychiatric and endocrine systems. The primary reason for the development of stress-induced disease is suppression of macrophage function, which plays a pivotal role in innate immunity. In fact, surgical stress has been shown to exacerbate opportunistic infections by significantly suppressing macrophage function. Conversely, administration of macrophage activating substances before surgery, such as tumor necrosis factor (TNF)-α or Picibanil (OK-432), has been shown to protect against macrophage suppression and the resulting exacerbation of infectious diseases, and against tumor metastasis in the lungs. Thus, if suppression of macrophage function by stress could be safely prevented by use of a macrophage activating substance, the detrimental side effects of stress could be reduced. Recently, we identified a lipopolysaccharide, IP-PA1, derived from Pantoea agglomerans, a symbiotic Gram-negative bacteria found in wheat and other food plants. Oral administration of IP-PA1 demonstrated macrophage activation (priming) and protective effects against infection, allergy and cancer, without any side-effects. In this review, the possibility of using IP-PA1 as a safe, macrophage activating substance for prevention of stress-induced impairments is discussed.

Nutrient availability and management in the rhizosphere: exploiting genotypic differences.

Crop nutrition is frequently inadequate as a result of the expansion of cropping into marginal lands, elevated crop yields placing increasing demands on soil nutrient reserves, and environmental and economic concerns about applying fertilizers. Plants exposed to nutrient deficiency activate a range of mechanisms that result in increased nutrient availability in the rhizosphere compared with the bulk soil. Plants may change their root morphology, increase the affinity of nutrient transporters in the plasma membrane and exude organic compounds (carboxylates, phenolics, carbohydrates, enzymes, etc.) and protons. Chemical changes in the rhizosphere result in altered abundance and composition of microbial communities. Nutrient-efficient genotypes are adapted to environments with low nutrient availability. Nutrient efficiency can be enhanced by targeted breeding through pyramiding efficiency mechanisms in a desirable genotype as well as by gene transfer and manipulation. Rhizosphere microorganisms influence nutrient availability; adding beneficial microorganisms may result in enhanced availability of nutrients to crops. Understanding the role of plant-microbe-soil interactions in governing nutrient availability in the rhizosphere will enhance the economic and environmental sustainability of crop production.

Siderophore production by fluorescent pseudomonads colonizing roots of certain crop plants.

Twelve fluorescent Pseudomonas isolates colonizing roots of four crop plants, chilli, cotton, groundnut and soybean, were examined for extracellular siderophore production in different media under iron deficient conditions. While all the organisms produced siderophores, they varied in the quantity of siderophores produced and in their preference to the medium. The siderophores were invariably hydroxamates (pyoverdine) of trihydroxamate type which formed bidentate ligands with Fe III ions.

Purification, characterisation and cDNA cloning of an antimicrobial peptide from Macadamia integrifolia.

An antimicrobial peptide with no significant amino acid sequence similarity to previously described peptides has been isolated from the nut kernels of Macadcamia integrifolia. The peptide, termed MiAMP1, is highly basic with an estimated pI of 10.1, a mass of 8.1 kDa and contains 76 amino acids including 6 cysteine residues. A cDNA clone containing the entire coding region corresponding to the peptide was obtained. The deduced amino acid sequence of the cDNA indicated a 26-amino-acid signal peptide at the N-terminus of the preprotein. Purified MiAMP1 inhibited the growth of a variety of fungal, oomycete and gram-positive bacterial phytopathogens in vitro. Some pathogens exhibited close to 100% inhibition in less than 1 microM peptide (5 microg/ml). Antimicrobial activity was diminished against most, but not all, microbes in the presence of calcium and potassium chloride salts (1 mM and 50 mM, respectively). MiAMP1 was active against bakers yeast, was inactive against Escherichia coli and was non-toxic to plant and mammalian cells. Analysis of genomic DNA indicated that MiAMP1 was encoded on a single copy gene containing no introns. The MiAMP1 gene may prove useful in genetic manipulations to increase disease resistance in transgenic plants.

Microhongos de la hierba mate (ilex paraguariensis st hil) / Microfungi of mate herb (ilex paraguariensis st hil)

Se estudió cuantitativa y cualitativamente la presencia de especies fúngicas mesófilicas, termotolerantes y termofilicas presentes en 10 muestras de yerba mate (ilex paraguariensis st. hil.) obtenidas de diversos locales comerciales de la capital federal del gran Buenos Aires. Mediante las técnicas de dilución en placa y cámara húmeda, se aislaron un total de 14 géneros y 15 especies. Los aislamientos más comunes fueron: aspergillus niger van tieghem ( en 110 por ciento de las muestras) y rhodotorula spp. (80 por ciento). Otros taxa menos frecuentes integraron los géneros aspergillus, penicillium, fusarium, paecilomyces y phaecoccomyces y algunas de ellos pueden considerarse potencialmente toxicogénicas. Cephaliophora tropica thaxter, melanospora sp., rhizomucor pusillus schipper y humicola grisea traaen, fueron unicamente recuperadas mediante la técnica de cultivo en cámara húmeda. El número de propágulos por gramo del producto analizado, osciló desde 2.700 a 50.000 ufc. para hongos mesofilicos y de 0-3000 ufc. para termotolerantes

Characterization and subcellular localization of tomato ringspot nepovirus putative movement protein.

Sequence comparison of the tomato ringspot nepovirus (TomRSV) genome with related viruses suggested that the region of the RNA-2-encoded polyprotein immediately upstream of the coat protein may be involved in the cell-to-cell movement of the virus (Rott et al., 1991, J. Gen. Virol. 72, 1505-1514). To further study the role of this portion of the genome, monoclonal antibodies against the putative movement protein were raised. Western blots of plant extracts allowed the detection of a viral nonstructural protein of M(r) 45K present only in TomRSV-infected tissues. Immunogold-labeling studies revealed that in Nicotiana clevelandii the putative movement protein was found only in infected cells immediately adjacent to the necrotic tissue, and that it was associated with tubular structures containing virus-like particles present in or near the cell wall. This provides further evidence that this protein is involved in the cell-to-cell movement of the virus and that this movement might take place via the formation of tubular structures.

Use of digoxigenin-labelled probes for detection and host-range studies of tomato yellow leaf curl geminivirus.

We studied the host range of tomato yellow leaf curl virus (TYLCV) in some agronomically important tomato species. Transmission tests with the natural vector Bemisia tabaci from tomato to sweet pepper, eggplant, cucumber, melon, zucchini and spinach showed that these species did not develop symptoms and did not support viral replication. These species therefore do not constitutive a reservoir of the virus and can be cultivated as alternatives to tomato in the most affected areas. For host-range studies, we used a quick and sensitive dot-blot assay employing non-radioactive DNA probes. This technique, developed for detecting TYLCV in plant extracts, is easily used for diagnosis. The sensitivity of this non-radioactive test was comparable to that of radiolabelled probes.

Mycoflora of anise and fennel seeds in Egypt.

Using four medium types (glucose-, cellulose-, 50% sucrose- and 10% NaCl-Czapek's agar), it was possible to isolate 15 fungal genera, 78 species and 6 varieties. The collective fungal spectrum varied from one medium to another where the highest number of species (57 species/1000 seeds) was obtained on glucose- and the lowest (31 species/1000 seeds) on 10% NaCl-Czapek's agar. Aspergillus, Penicillium, and sometimes Rhizopus and Chaetomium were the most common genera on the different medium types. The most common fungal species especially on glucose- and cellulose-Czapek's agar were, Aspergillus niger, A. flavus, A. ochraceus, A. fumigatus, A. flavus var. columnaris, Penicillium chrysogenum, Rhizopus stolonifer and Chaetomium globosum. Members of A. glaucus group were more frequently recovered on 10% NaCl- and 50% sucrose-Czapek's agar.