DGGE and real‐time PCR analysis of lactic acid bacteria in bacterial communities of the phyllosphere of lettuce

Zwielehner, Jutta; Handschur, Michael; Michaelsen, Astrid; Irez, Selen; Demel, Michael A.; Denner, Ewald B. M.; Haslberger, Alexander G.

doi:10.1002/mnfr.200700158

Cited by 44 publications

(37 citation statements)

References 35 publications

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“…Mycobacteria (predominantly M. avium) were isolated from 46 samples of salads, leak, lettuce, mushrooms, and other vegetables as well as apple juice and twenty nine isolates were tested (Argueta et al 2000). Zwielehner et al (2008) studied the microbial communities present in the phylosphere of lettuce leaves. After denaturing gradient gel electrophoresis and sequencing analyses, sequences from the genus Mycobacterium were found on leaves as well as soil samples.…”

Section: Mycobacteria In Soilmentioning

confidence: 99%

Mycobacteria in water, soil, plants and air: a review

Hruška¹,

Kaevska²

2012

Vet. Med.

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Amazingly, despite the 24 143 papers on mycobacteria, indexed in the Web of Science database during the last six years, published by 67 008 authors from 13 128 organizations located in 166 countries or territories, internationally accepted legal directives on how to control the public health risk associated with environmental mycobacteria have yet to be developed. Mycobacteria are human and animal pathogens, causing not only tuberculosis and leprosy, but mycobacterioses of skin, soft tissues and lung. Due to their cell wall composition and their adaptability mycobacteria can survive in different habitats for years. Their immunomodulatory ability has been recognised for more than 50 years and hundreds of papers published during the last two decades have demonstrated that small chemical products derived from mycobacterial cells participate in inflammatory pathways involved the pathogenesis of important human diseases like Crohn's disease, asthma, type 1 diabetes mellitus, psoriasis, arthrosis, Blau syndrom, sarcoidosis, autism etc. Mycobacteria can influence inflammatory pathways not only as live organisms, but also by means of components derived from dead cells. Pasteurisation or cooking does not affect this ability. Hence, how many mycobacterial cells are ingested, what factors play a role concurrently, and how long the harmful effect persists become important questions. This paper presents only a short review based on selected papers about mycobacteria in water, soil, plants and air with the aim of attracting attention to this significant global problem and of making the first steps towards protection of people. Selected bibliographic references of published data from 2007 to 2012 are presented in easy-to-navigate tables.Keywords: Mycobacterium; water; soil; plant; vegetables; air; biofilm; sediment; determination; zoonoses; food safety List of abbreviations CFU = colony forming units; DGGE = denaturation gradient gel electrophoresis; IMS = immuno-magnetic separation; IS = insertion sequence; ITS = intergenic transcribed spacer; MAC = Mycobacterium avium complex; MAP/Map = Mycobacterium avium subsp. paratuberculosis; MTC = Mycobacterium tuberculosis complex; MWF = metal working fluids; nPCR = nested PCR; NTM = non-tuberculous mycobacteria; PCR = polymerase chain reaction; PPM = potentially pathogenic mycobacteria; qPCR = quantitative real time PCR; RFLP = restriction fragment length polymorphism

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Section: Mycobacteria In Soilmentioning

confidence: 99%

Mycobacteria in water, soil, plants and air: a review

Hruška¹,

Kaevska²

2012

Vet. Med.

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“…Bacterial communities associated with leafy greens have already been described by several culture-independent studies. "First-generation" molecular techniques have been used to describe variation in community structure in the context of leaf surface properties and microbial interactions (5), seasonal variation in the community structure (6), and monitoring of bacterial communities in the food chain (7). The introduction of culture-independent methods, in particular, microbial profiling using high-throughput sequencing to study microorganisms, has revealed more complexity and diversity of the phyllosphere microbiota and has dramatically changed the landscape of microbial ecology (5,(7)(8)(9)(10).…”

mentioning

confidence: 99%

Bacterial Communities Associated with Surfaces of Leafy Greens: Shift in Composition and Decrease in Richness over Time

Dees

Lysøe

Nordskog

et al. 2015

Appl Environ Microbiol

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The phyllosphere is colonized by a wide variety of bacteria and fungi; it harbors epiphytes, as well as plant-pathogenic bacteria and even human pathogens. However, little is known about how the bacterial community composition on leafy greens develops over time. The bacterial community of the leafy-green phyllosphere obtained from two plantings of rocket salad (Diplotaxis tenuifolia) and three plantings of lettuce (Lactuca sativa) at two farms in Norway were profiled by an Illumina MiSeq-based approach. We found that the bacterial richness of the L. sativa samples was significantly greater shortly (3 weeks) after planting than at harvest (5 to 7 weeks after planting) for plantings 1 and 3 at both farms. For the second planting, the bacterial diversity remained consistent at the two sites. This suggests that the effect on bacterial colonization of leaves, at least in part must, be seasonally driven rather than driven solely by leaf maturity. The distribution of phyllosphere communities varied between D. tenuifolia and L. sativa at harvest. The variability between these species at the same location suggests that the leaf-dwelling bacteria are not only passive inhabitants but interact with the host, which shapes niches favoring the growth of particular taxa. This work contributes to our understanding of host plant-specific microbial community structures and shows how these communities change throughout plant development.T he phyllosphere is a habitat on the surface of plant leaves colonized by a variety of bacteria, yeasts, and fungi (1). It harbors epiphytes, as well as plant-pathogenic bacteria and even human pathogens. Microbial populations on leaf surfaces are highly influenced by rapid fluctuations in UV radiation, temperature, and humidity and are restricted by limited access to nutrients (1, 2). Resident bacteria on leaves can have neutral, negative, or positive influences on their host plants by serving as pathogens, preventing leaf colonization by pathogens, or acting as growth promoters (3). Traditionally, phyllosphere bacteria have been characterized by using culture-based approaches and much of the work on produce-associated bacteria has focused on a small number of pathogenic species. Culture-based methods will not include bacteria that are not able to grow on standard artificial media or are slow growing. This limits our understanding of the phyllosphere microbial community's ecology, genetics, and physiology (4). Bacterial communities associated with leafy greens have already been described by several culture-independent studies. "First-generation" molecular techniques have been used to describe variation in community structure in the context of leaf surface properties and microbial interactions (5), seasonal variation in the community structure (6), and monitoring of bacterial communities in the food chain (7). The introduction of culture-independent methods, in particular, microbial profiling using high-throughput sequencing to study microorganisms, has revealed more complexity and diversity of the...

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“…Different lifestyle factors and diets were shown to have a significant impact on the faecal microbiota [2,3] .…”

Section: Introductionmentioning

confidence: 99%

Characterization of Bacteria, Clostridia and<i> Bacteroides</i> in Faeces of Vegetarians Using qPCR and PCR-DGGE Fingerprinting

et al. 2009

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Background/Aims: This study aimed to investigate the quantitative and qualitative changes of bacteria, Bacteroides, Bifidobacterium and Clostridium cluster IV in faecal microbiota associated with a vegetarian diet. Methods: Bacterial abundances were measured in faecal samples of 15 vegetarians and 14 omnivores using quantitative PCR. Diversity was assessed with PCR-DGGE fingerprinting, principal component analysis (PCA) and Shannon diversity index. Results: Vegetarians had a 12% higher abundance of bacterial DNA than omnivores, a tendency for less Clostridium cluster IV (31.86 ± 17.00%; 36.64 ± 14.22%) and higher abundance of Bacteroides (23.93 ± 10.35%; 21.26 ± 8.05%), which were not significant due to high interindividual variations. PCA suggested a grouping of bacteria and members of Clostridium cluster IV. Two bands appeared significantly more frequently in omnivores than in vegetarians (p < 0.005 and p < 0.022). One was identified as Faecalibacterium sp. and the other was 97.9% similar to the uncultured gut bacteriumDQ793301. Conclusions: A vegetarian diet affects the intestinal microbiota, especially by decreasing the amount and changing the diversity of Clostridium cluster IV. It remains to be determined how these shifts might affect the host metabolism and disease risks.

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DGGE and real‐time PCR analysis of lactic acid bacteria in bacterial communities of the phyllosphere of lettuce

Cited by 44 publications

References 35 publications

Mycobacteria in water, soil, plants and air: a review

Mycobacteria in water, soil, plants and air: a review

Bacterial Communities Associated with Surfaces of Leafy Greens: Shift in Composition and Decrease in Richness over Time

Characterization of Bacteria, Clostridia and<i> Bacteroides</i> in Faeces of Vegetarians Using qPCR and PCR-DGGE Fingerprinting

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