Responses of Escherichia coli and Listeria monocytogenes to ozone treatment on non-host tomato: Efficacy of intervention and evidence of induced acclimation.

Because of the continuous rise of foodborne illnesses caused by the consumption of raw fruits and vegetables, effective post-harvest anti-microbial strategies are necessary. The aim of this study was to evaluate the anti-microbial efficacy of ozone (O3) against two common causes of fresh produce contamination, the Gram-negative Escherichia coli O157:H7 and Gram-positive Listeria monocytogenes, and to relate its effects to potential mechanisms of xenobiosis by transcriptional network modeling. The study on non-host tomato environment correlated the dose × time aspects of xenobiosis by examining the correlation between bacterial survival in terms of log-reduction and defense responses at the level of gene expression. In E. coli, low (1 µg O3/g of fruit) and moderate (2 µg O3/g of fruit) doses caused insignificant reduction in survival, while high dose (3 µg/g of fruit) caused significant reduction in survival in a time-dependent manner. In L. monocytogenes, moderate dose caused significant reduction even with short-duration exposure. Distinct responses to O3 xenobiosis between E. coli and L. monocytogenes are likely related to differences in membrane and cytoplasmic structure and components. Transcriptome profiling by RNA-Seq showed that primary defenses in E. coli were attenuated after exposure to a low dose, while the responses at moderate dose were characterized by massive upregulation of pathogenesis and stress-related genes, which implied the activation of defense responses. More genes were downregulated during the first hour at high dose, with a large number of such genes getting significantly upregulated after 2 hr and 3 hr. This trend suggests that prolonged exposure led to potential adaptation. In contrast, massive downregulation of genes was observed in L. monocytogenes regardless of dose and exposure duration, implying a mechanism of defense distinct from that of E. coli. The nature of bacterial responses revealed by this study should guide the selection of xenobiotic agents for eliminating bacterial contamination on fresh produce without overlooking the potential risks of adaptation.

Antibacterial Mechanism of Liangguoan against Staphylococcus aureus and Escherichia coli.

This study examined the antibacterial activity of the biological pesticide Liangguoan against Staphylococcus aureus and Escherichia coli as a potential replacement for chemical pesticide use in the fruit and vegetable industry. We measured the minimum inhibitory concentration and observed the changes in bacterial morphology, mortality, conductivity, nucleic acid content, and ATP content in response to the bactericide. The minimum inhibitory concentration of Liangguoan was 20 mg/mL for S. aureus and 40 mg/mL for E. coli. After treatment with Liangguoan, the mortality rates of S. aureus and E. coli reached 78.3% and 63.7%, respectively. We observed that the cells were scattered and that the cell morphology was altered in that the cells shortened. The interconnection effect and ATP content decreased, whereas cell conductivity and the nucleic acid content increased. In summary, Liangguoan inhibited S. aureus and E. coli by destroying their cell structure and disrupting their metabolism.

Anti-listerial properties of chemical constituents of Eruca sativa (rocket salad): From industrial observation to in vitro activity.

The frequency of foodborne outbreaks epidemiologically associated with Listeria monocytogenes in fresh produce has increased in recent years. Although L. monocytogenes may be transferred from the environment to vegetables during farming, contamination of food products most commonly occurs in food processing facilities, where L. monocytogenes has the ability to establish and persist on processing equipment. The current study was undertaken to collect data on the occurrence of L. monocytogenes and the identity of the endogenous microbiota in a fresh produce processing facility, for which information has remained scarce. L. monocytogenes was not detected in the facility. Experiments simulating conditions in the processing environment were performed, including examination of bacterial growth in nutrients based on vegetables (salad juice) compared to in other types of nutrients (fish, meat). Results showed that the endogenous microbiota (dominated by Pseudomonas) grew well in iceberg lettuce and rocket salad juice at low temperatures, while growth inhibition of L. monocytogenes was observed, particularly in rocket salad juice. The anti-listerial activity in rocket salad juice was retained in a polar chromatographic fraction containing several metabolites. Characterization of this active fraction, using LC-MS/MS, led to identification of 19 compounds including nucleosides and amino acids. Further work is necessary to determine the molecular mechanism responsible for the inhibitory activity of rocket salad constituents. The study nevertheless suggests that the available nutrients, as well as a low temperature (3 °C) and the in-house bacterial flora, may influence the prevalence of L. monocytogenes in fresh produce processing facilities.

Fermentation of African nightshade leaves with lactic acid bacterial starter cultures.

The interest in the consumption of African indigenous leafy vegetables increased in African countries, e.g. Kenya, within the last years. One example of African indigenous leafy vegetables is African nightshade (Solanum scabrum) which is nutritious, rich in proteins and micronutrients and therefore could contribute to a healthy diet. African nightshade has several agricultural advantages. However, the most important disadvantage is the fast perishability which leads to enormous post-harvest losses. In this study, we investigated the fermentation of African nightshade as a post-harvest processing method to reduce post-harvest losses. The two lactic acid bacterial starter strains Lactiplantibacillus plantarum BFE 5092 and Limosilactobacillus fermentum BFE 6620 were used to inoculate fermentations of African nightshade leaves with initial counts of 106-107 cfu/ml. Uninoculated controls were conducted for each fermentation trial. Fermentations were performed both in Kenya and in Germany. The success of the inoculated starter cultures was proven by the measurement of pH values and determination of lactic acid concentration. Lactobacilli strains dominated the microbiota of the starter inoculated samples in contrast to the non-inoculated controls. This was supported by classical culture-dependent plating on different microbiological media as well as by the culture-independent molecular biological methods denaturing gradient gel electrophoresis and 16S rRNA gene high-throughput amplicon sequencing. We could demonstrate that the use of the selected starter cultures for fermentation of African nightshade leaves led to controlled and reliable fermentations with quick acidification. Thus, controlled fermentation with appropriate starter cultures is a promising method for post-harvest treatment of African nightshade leaves.

Occurrence and diversity of ciliated protozoa in the most commonly consumed leafy greens in Brazil: a qualitative study / Ocorrência e diversidade de protozoários ciliados nos vegetais folhosos mais consumidos no Brasil: um estudo qualitativo

Ciliated protozoa are an ubiquitous group of eukaryotes that have been poorly reported on leafy greens. The present study aimed to verify the occurrence and diversity of ciliated protozoa in the leaves and roots of three of the most commonly consumed leafy greens in Brazil – lettuce, rocket and coriander. The vegetable samples were washed by manual agitation (3 minutes) in two different media (mineral water and Page ́s Amoeba Saline solution). After washing, the contents were incubated in Petri dishes and aliquots were removed for microscopic identification and in vivo observation. A total of 21 ciliated protozoa species were found, most of which were bacterivorous. Leafy greens have commonly been associated with foodborne outbreaks and ciliated protozoa, which although they are not a Public Health concern, can act as “Trojan Horses” harboring bacteria, viruses and other protozoa cysts and oocysts and can suggest a new route towards microbiological quality related to the food chain. This is the first report of ciliated protozoa on leafy greens consumed in Brazil.

Os protozoários ciliados constituem um grupo onipresente de eucariotos pouco relatados em vegetais folhosos. O presente estudo teve como objetivo verificar a ocorrência e a diversidade de protozoários ciliados nas folhas e raízes de três dos vegetais folhosos mais consumidos no Brasil - alface, rúcula e coentro. As amostras de vegetais foram lavadas por agitação manual (3 minutos) em dois meios diferentes (água mineral e Solução Salina para Ameba). Após a lavagem, o conteúdo foi incubado em placas de Petri e alíquotas foram retiradas para identificação microscópica e observação in vivo dos organismos. Um total de 21 espécies de protozoários ciliados foi encontrado, a maioria das quais era bacterívora. Tais hortaliças têm sido comumente associadas a surtos transmitidos por alimentos e os protozoários ciliados que, embora não sejam um problema de saúde pública, podem atuar como “Cavalos de Tróia” ao abrigar bactérias, vírus além de cistos e oocistos de protozoários o que pode sugerir uma nova rota para a avaliação da qualidade microbiológica relacionada à cadeia alimentar. Este é o primeiro relato de protozoários ciliados em folhas verdes consumidas no Brasil.

Macrophage activation by edible mushrooms is due to the collaborative interaction of toll-like receptor agonists and dectin-1b activating beta glucans derived from colonizing microorganisms.

Research supports the theory that the microbiome of plants and mushrooms produce potent activators of pathogen recognition receptors which are principal contributors to the stimulation of macrophages. We have previously reported that the in vitro macrophage stimulatory activity of water-soluble extracts from 13 different types of edible mushrooms is predominantly due to bacterial components originating from the naturally occurring bacterial communities within these materials. The purpose of the current study was to further investigate the bacterial-dependent activity of the water-soluble extracts and assess whether these 13 types of mushrooms contain water-insoluble beta glucans that activate the dectin-1b signaling pathway. Activity of the water-soluble extracts was predominantly due to Toll-like receptor 2 (TLR2) and TLR4 agonists. For dectin-1b-dependent activity (indicative of water-insoluble beta glucans), culinary mushrooms (Agaricus bisporus varieties) were essentially inactive, whereas most of the medicinal mushrooms (Lentinula edodes, Grifola frondosa, Hypsizygus marmoreus varieties, Flammulina velutipes) exhibited potent activation. A. bisporus samples with no detectable dectin-1b-dependent activity had yeast colony forming units that were 687 times lower than L. edodes exhibiting high activity, indicating that the active insoluble beta glucans are derived from colonizing yeast. In addition, co-stimulation of macrophages with the TLR agonists and insoluble beta glucan was found to result in a synergistic enhancement of in vitro cytokine production. Taken together, these findings indicate that the in vitro macrophage activating potential of edible mushrooms is due to the collaborative interaction of water-soluble TLR agonists (derived from colonizing bacteria) and water-insoluble beta glucans (derived from colonizing yeast).

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.

Plant species affects establishment of Escherichia coli O157:H7 gfp+ on leafy vegetables.

AIMS: Greenhouse trials were conducted with different cultivars of baby leaf spinach, rocket and Swiss chard and inoculation of Escherichia coli O157:H7 gfp+, to determine whether plant species and cultivar have an impact on the establishment of this strain. METHODS AND RESULTS: Three cultivars each of spinach, rocket and Swiss chard were spray inoculated with E. coli O157:H7 gfp+ at doses of log 7 CFU per ml. Due to the different lengths of growing period spinach and Swiss chard were spray inoculated three times and rocket five times, with final inoculation performed 3 days prior to harvest. After a growing period of 26-33 days, E. coli O157:H7 gfp+ was recovered from the leaf surface in mean populations between log 1 and 6 CFU per gram. The lowest occurrence of E. coli O157:H7 gfp+ was found on rocket leaves and the highest on spinach. There was no significant difference in the establishment of E. coli O157:H7 gfp+ between cultivars, but there were differences between plant species. Indigenous phyllosphere bacteria were pure cultured and identified with 16S rRNA gene sequencing. CONCLUSIONS: Despite the same high inoculation dose of E. coli O157:H7 gfp+ on leaves, the establishment rate differed between plant species. However, plant cultivar did not affect establishment. Pantoea agglomerans dominated the identified bacterial isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: As previous studies are inconclusive on choice of model plant species and cultivar, we studied whether plant species or cultivar determines the fate of E. coli O157:H7 gfp+ on leafy vegetables. The findings indicate that plant species is a key determinant in the establishment of E. coli O157:H7 gfp+.

Influence of Plant Species, Tissue Type, and Temperature on the Capacity of Shiga-Toxigenic Escherichia coli To Colonize, Grow, and Be Internalized by Plants.

Contamination of fresh produce with pathogenic Escherichia coli, including Shiga-toxigenic E. coli (STEC), represents a serious risk to human health. Colonization is governed by multiple bacterial and plant factors that can impact the probability and suitability of bacterial growth. Thus, we aimed to determine whether the growth potential of STEC for plants associated with foodborne outbreaks (two leafy vegetables and two sprouted seed species) is predictive of the colonization of living plants, as assessed from growth kinetics and biofilm formation in plant extracts. The fitness of STEC isolates was compared to that of environmental E. coli isolates at temperatures relevant to plant growth. Growth kinetics in plant extracts varied in a plant-dependent and isolate-dependent manner for all isolates, with spinach leaf lysates supporting the highest rates of growth. Spinach extracts also supported the highest levels of biofilm formation. Saccharides were identified to be the major driver of bacterial growth, although no single metabolite could be correlated with growth kinetics. The highest level of in planta colonization occurred on alfalfa sprouts, though internalization was 10 times more prevalent in the leafy vegetables than in sprouted seeds. Marked differences in in planta growth meant that the growth potential of STEC could be inferred only for sprouted seeds. In contrast, biofilm formation in extracts related to spinach colonization. Overall, the capacity of E. coli to colonize, grow, and be internalized within plants or plant-derived matrices was influenced by the isolate type, plant species, plant tissue type, and temperature, complicating any straightforward relationship between in vitro and in planta behaviors.IMPORTANCE Fresh produce is an important vehicle for STEC transmission, and experimental evidence shows that STEC can colonize plants as secondary hosts, but differences in the capacity to colonize occur between different plant species and tissues. Therefore, an understanding of the impact that these plant factors have on the ability of STEC to grow and establish is required for food safety considerations and risk assessment. Here, we determined whether growth and the ability of STEC to form biofilms in plant extracts could be related to specific plant metabolites or could predict the ability of the bacteria to colonize living plants. Growth rates for sprouted seeds (alfalfa and fenugreek) but not those for leafy vegetables (lettuce and spinach) exhibited a positive relationship between plant extracts and living plants. Therefore, the detailed variations at the level of the bacterial isolate, plant species, and tissue type all need to be considered in risk assessment.

Fungicide-driven alterations in azole-resistant Aspergillus fumigatus are related to vegetable crops in Colombia, South America.

Aspergillus fumigatus resistant to azole as first-line therapy has been reported in azole-naïve patients. This worldwide resistance phenomenon has been linked to fungicide-driven alterations in the cyp51A gene and its promoter region (such as TR34/L98H and TR46/Y121F/T289A). Azole-resistant A. fumigatus related to the use of triazole fungicides in flower fields was recently reported In Colombia. The purpose of this study was to investigate the presence of azole-resistant A. fumigatus in soil samples from vegetable crops such as carrots, potatoes, maize, strawberries, and pea, and from prepared farming land surrounding the city of Bogotá. Species identification was based on sequencing of the ß-tubulin and calmodulin genes. All A. fumigatus strains were screened for azole resistance on agar supplemented with itraconazole or voriconazole. Among the 60 soil samples, 34 (56.6%) were positive for A. fumigatus and 15 samples exhibited strains (n = 18) that grew on agar supplemented with itraconazole or voriconazole. Triazole-resistant strains were isolated from soil samples associated with carrot, potato, maize, and pea crops. Sequencing of the cyp51A gene and its promoter region indicated polymorphism, mainly with the presence of TR46/Y121F/T289A (n = 8), TR34/L98H, and TR53. Eight resistant isolates exhibited cyp51A wild type without alterations in the promoter region. Our study showed evidence of dissemination of azole-resistant A. fumigatus, with high genetic diversity, in vegetable crops in Colombia. These data underline the need to determine the prevalence of azole resistance in A. fumigatus in clinical and environmental settings for other regions of Colombia as well as Latin America.

Endophytic bacterial communities in peels and pulp of five root vegetables.

Plants contain endophytic bacteria, whose communities both influence plant growth and can be an important source of probiotics. Here we used deep sequencing of a 16S rRNA gene fragment and bacterial cultivation to independently characterize the microbiomes of five plant species from divergent taxonomic orders-potato (Solanum tuberosum), carrot (Daucus sativus), beet (Beta vulgaris), neep (Brassica napus spp. napobrassica), and topinambur (Helianthus tuberosus). We found that both species richness and diversity tend to be higher in the peel, where Alphaproteobacteria and Actinobacteria dominate, while Gammaproteobacteria and Firmicutes dominate in the pulp. A statistical analysis revealed that the main characteristic features of the microbiomes of plant species originate from the peel microbiomes. Topinambur pulp displayed an interesting characteristic feature: it contained up to 108 CFUs of lactic acid bacteria, suggesting its use as a source of probiotic bacteria. We also detected Listeria sp., in topinambur pulps, however, the 16S rRNA gene fragment is unable to distinguish between pathogenic versus non-pathogenic species, so the evaluation of this potential health risk is left to a future study.

Acidogenic properties of carbohydrate-rich wasted potato and microbial community analysis: Effect of pH.

Vegetable waste is one of the major organic solid residues available for sustainable biogas production. The aim of this study was to investigate the possibility and optimal controlling strategy for acidogenic fermentation of wasted potato (WP). Three leaching bed reactors (LBRs) were operated at various pH values (6.0, 7.0 and 8.0) with an organic loading rate (OLR) of 6.7 g volatile solid/(L·d) and hydraulic retention time of 6 d. Butyric acid-type fermentation with butyric acid as predominant volatile fatty acid (VFA) was observed with a concentration and proportion (of total VFAs) of butyric acid, which were 7.8 g/L, 49.7 % and 9.6 g/L and 52.2 % at pH 6.0 and 7.0, respectively. Conversely, at pH 8.0, mixed acid-type fermentation was observed with acetic and butyric acid as the major VFAs. Control experiment without pH manipulation didn't perform well in VFAs production at first 6 days and then VFAs concentration increased as pH value was adjusted to 8. It was indicated that the inhibition was caused by high undissociated VFAs concentration due to low pH and the VFAs production could be improved through pH control strategy to regulate the undissociated VFAs concentration. According to the bacterial analysis, the microbial community was diverse and Firmicutes were the most important bacteria at different pH conditions. Therefore, the results suggested that a process of pH control might be feasible for stable and efficient acidogenic fermentation.

Occurrence and species diversity of human-pathogenic Mucorales in commercial food-stuffs purchased in Paris area.

Mucormycoses are life-threatening fungal diseases that affect a variety of patients including those with diabetes mellitus or hematological malignancies. The responsible agents, the Mucorales, are opportunistic pathogens originating from the environment such as soil or decaying organic matter. The aim of the present study was to assess the prevalence and diversity of human-pathogenic species of Mucorales in commercially available foodstuffs in France. All food samples were purchased from January 2014 to May 2015 in France. A total of 159 dried food samples including spices and herbs (n = 68), herbal tea (n = 19), cereals (n = 19), vegetables (n = 14), and other foodstuffs (n = 39) were analyzed. Each strain of Mucorales was identified phenotypically, and molecular identification was performed by ITS sequencing. From the 28 (17.6%) samples that were culture-positive for Mucorales, 30 isolates were recovered. Among the isolates, 13 were identified as Rhizopus arrhizus var. arrhizus, 10 R. arrhizus var. delemar, two Rhizopus microsporus, one Lichtheimia corymbifera, three Lichtheimia ramosa, and one Syncephalastrum racemosum. Culture-positive samples originated from different countries (Europe, Asia) and brands. The samples most frequently contaminated by Mucorales were spices and herbs (19/68, 27.9%), followed by herbal tea (2/19, 10.5%), cereals (2/19, 10.5%), other food products (5/39, 12.8%). The present study showed that human-pathogenic Mucorales were frequently recovered from commercially available foodstuffs in France with a large diversity of species. The potential danger represented by Mucorales present in food for immunocompromised patients should be further analyzed.

High Clostridium difficile contamination rates of domestic and imported potatoes compared to some other vegetables in Slovenia.

Clostridium difficile, recently reclassified to Clostridioides difficile, is among most important causes of intestinal infections in humans. Zoonotic potential and foodborne transmissions are considered to be partially involved in C. difficile spread. Here we report prevalence of C. difficile in 142 retail and 12 homegrown vegetables in Slovenia between years 2014 and 2017. The overall prevalence of C. difficile on vegetables was 18,2% (28/154). A total of 115 isolates were obtained which belonged to 25 PCR ribotypes. Ten of those were toxigenic and PCR ribotype 014/020 was the most prevalent. Most of 25 determined PCR ribotypes were previously reported in humans, animals, soil or water in Slovenia. Among tested vegetables, potatoes had the highest positivity rate (28,0% vs. 6,7% and 9,4% for ginger and leaf vegetables). Altogether 66,7% of C. difficile positive potato samples were imported from 12 different countries of three different continents. The origin of contamination could be any point between production and retail store, however, our results suggest a possibility that potatoes represent a transnational and transcontinental way of C. difficile transmissions.

Epidemiology of Pseudomonas aeruginosa in agricultural areas.

A prevailing opinion is that the strains of Pseudomonas aeruginosa that infects both plants and humans are two separate species. This study strongly disputes that notion until the modern molecular technology proves otherwise. This paper examines a spectrum of strains occurring in nature, their habitats, dissemination, their relationship to clinical strains, and the environmental conditions that favor their colonization of plants. The isolates were obtained from clinical specimens, plants, soil, and water. The identity of these strains was confirmed using pyocin typing and biochemical assays. The data reveal that agricultural soils, potted ornamental plants, hoses, fountains, and faucets frequently harbored P. aeruginosa. However, it was not commonly found in semi-arid areas, suggesting that moisture and high humidity is necessary for colonization and survival. Though found in soil, P. aeruginosa was seldom isolated on edible plant parts. The pathogenicity of various strains on plants was tested by inoculating vegetables, lettuce slices (Lactuca sativa L. "Great Lakes"), celery stalks (Apium graveolens L. var. Dulce], potato tuber slices (Solanum tuberosum L. "Whiterose"), tomato (Lycopersicon esculentum L. Mill), cucumber (Cucumis sativus L.), rutabaga (Brassica campestris L.), and carrot (Daucus carota L. var sativa). There was considerable variation in the strains' ability to cause rot, but no difference was observed between clinical isolates and others from agricultural fields, water, and soil. Two of the clinical isolates from burn patients, P. aeruginosa PA13 and PA14, exhibited the greatest virulence in causing rot in all the plants that were tested, especially on cucumber, lettuce, potato, and tomato. The study discusses how closely the epidemiology of P. aeruginosa relates to many plant pathogens, and the ability of human isolates to colonize plants and food material under favorable conditions. The biochemical and phenotypic similarity among strains from the clinical and agricultural material is strongly indicative that they are the same species and that plants and soil are natural reservoirs for P. aeruginosa.

Lactobacillus curvatus HY7601 and Lactobacillus plantarum KY1032 Cell Extracts Inhibit Adipogenesis in 3T3-L1 and HepG2 Cells.

Some lactic acid bacteria (LAB) and their cellular components have antiobesity effects. In this study, we evaluated the antiadipogenic effects of a mixture of two LAB-Lactobacillus curvatus HY7601 and Lactobacillus plantarum KY1032-using 3T3-L1 preadipocytes and HepG2 hepatocarcinoma cells. 3T3-L1 cells treated with a 1:1 ratio of HY7601 and KY1032 during differentiation showed reduced lipid accumulation by Oil Red O staining, as well as decreased leptin secretion and mRNA expression of peroxisome proliferator-activated receptor-γ and CCAAT/enhancer binding protein-α. HY7601 and KY1032 treatment also suppressed mitochondrial biogenesis and inhibited the expression of genes encoding mitochondrial transcription factors, as well as those related to fatty acid synthesis in HepG2 cells. The antiadipogenic effects of LAB were associated with the cell membrane fraction. These results demonstrate that a mixture of two LAB (HY7601 and KY1032) inhibits adipogenesis in preadipocytes and liver cells and is a potential therapeutic strategy for the treatment of obesity.

Microbial quality of organic and conventional vegetables from Polish farms.

Microbiological analyses of lettuce, radish, carrot and beetroot were conducted to determine the effect of production system (organic and conventional) on the microbial quality of vegetables in Poland. During 2010-2014 growing seasons, 600 organic and 372 conventional samples were collected from certified farms. The vegetables were analyzed for aerobic mesophilic bacteria, yeasts and molds, Enterobacteriaceae, coliforms and Escherichia coli according to Polish standards. The farmer's survey was conducted to collect information on farm management practices. The index (from 0 - no risk to 4 - high risk) of potential contamination of the produce by human pathogens, related to fertilization system was developed. The mesophilic bacteria, yeasts and molds, coliforms and Enterobacteriaceae numbers for the radish and carrot were similar for organic and conventional cultivation systems (mesophilic bacteria 7.0 log10 cfu g-1 and 6.6 log10 cfu g-1; yeasts and molds 5.1 and 4.8; coliforms 1.3 and 1.5; Enterobacteriaceae 2.1 and 2.3 for radish and carrot respectively). Organic lettuce harbored significantly more bacteria than conventional (mesophilic 6.7 log10 cfu g-1 and 6.4 log10 cfu g-1, coliforms 1.8 and 1.4; Enterobacteriaceae 2.5 and 1.9 for organic and conventional respectively). Organic beetroot contained higher number of yeasts and molds (5.1 log10 cfu g-1) and Enterobacteriaceae (2.9 log10 cfu g-1) than conventional (4.9 and 2.5 log10 cfu g-1). The vegetables from organic farms showed significantly higher load of E. coli (on average 0.42 log10 cfu g-1) than conventionally cultivated vegetables (in average 0.05 log10 cfu g-1). The index 0-4 of potential risk of produce contamination by human pathogens was created according to fertilization practices in both farm types. Its value increased with enhanced contribution of manures and other animal wastes. In organic production the main fertilization practice was application of animal manures, composted and not composted. A popular practice was also top dressing of growing plants with fermented plant extracts, sometimes enriched with dungwater. In conventional farming system mineral fertilization was the main source of the vegetable nutrition. Therefore, organic produce indicated higher index of contamination risk (2-4) than conventional vegetables (1-2). High indexes were positively associated with higher number of E. coli. It was found that fertilization system practiced in organic farms may deteriorate sanitary quality of the produce.

Screening of antifungal activity of 12 essential oils against eight pathogenic fungi of vegetables and mushroom.

The antifungal properties of 12 Eos, that is, Syzygium aromaticum, Pelargonium graveolens, Lavandula angustifolia, Cupresus sempervirens, Mentha piperita, Santolina chamaecyparissus, Citrus sinensis, Pogostemon patchouli, Thymus mastichina, Thymus vulgaris, Eucalyptus globulus and Rosmarinus officinalis, were screened. The influence of five doses of each EOs was tested against Botrytis cinerea, Sclerotinia sclerotiorum, Fusarium oxysporum, Phytophthora parasitica, Pythium aphanidermatum, Alternaria brassicae, Cladobotryum mycophilum and Trichoderma aggressivum f.sp. europaeum using disc-diffusion method. The mycelial growth inhibition and ED50 were calculated. The chemical analysis of the EOs was analysed using gas chromatography-mass spectroscopy. A total of 58 compounds were identified in the 12 EOs. All essential oils (EOs) analysed showed antifungal activity against the test pathogens in the range of 5·32-100%. The inhibitory effect of oils showed dose-dependent activity on the tested fungus. Based on the ED50 values, clove, rose geranium, peppermint and patchouli were the most effective. This study warrants further research into the practical use of EOs for the control of important myco- and phytopathogens in intensive horticulture. SIGNIFICANCE AND IMPACT OF THE STUDY: Due to the serious damage caused by fungal pathogens of vegetables and mushrooms, it is necessary to search for integrated strategies of disease control. This study provides relevant information about the effects of 12 essential oils (EOs) against eight pathogens of agricultural interest, included mycopathogens with emphasis on the possible future application of the EOs as alternative antifungal agents.

Lactobacillus curvatus KFP419 and Leuconostoc mesenteroides subsp. mesenteroides KDK411 Isolated from Kimchi Ameliorate Hypercholesterolemia in Rats.

Western-style diets increase the risk for cardiovascular diseases. It is suggested that the risk could be prevented by lowering cholesterol concentrations in blood. In the present study, hypocholesterolemic effects of the probiotics isolated from kimchi (Lactobacillus curvatus KFP419, Leuconostoc paramesenteroides KJP421, and Leuconostoc mesenteroide subsp. mesenteroides KDK411) were investigated in hypercholesterolemia-induced rats. There was no difference in growth performance between the rats fed high cholesterol diet (HCD) and normal diet (ND). However, blood total cholesterol, low-density lipoprotein cholesterol, and hepatic cholesterol were elevated by the HCD compared to ND, and those concentrations were decreased by dietary supplementation of KFP419 and KDK411. It was concomitant with an increase in fecal excretion of neutral sterols (cholesterol, coprostanol, and coprostanone) in the rats fed HCD compared to ND and was even greater with KDK411 supplementation. These findings indicate that probiotics L. curvatus KFP419 and L. mesenteroide subsp. mesenteroides KDK411 isolated from kimchi ameliorate hypercholesterolemia in rats by assimilating and excreting cholesterol in feces.