Deciphering the microbiome shift during fermentation of medicinal plants.

The importance of the human-microbiome relationship for positive health outcomes has become more apparent over the last decade. Influencing the gut microbiome via modification of diet represents a possibility of maintaining a healthy gut flora. Fermented food and lactic acid bacteria (LAB) display a preventive way to inhibit microbial dysbioses and diseases, but their ecology on plants is poorly understood. We characterized the microbiome of medicinal plants (Matricaria chamomilla L. and Calendula officinalis L.) using 16S rRNA gene profiling from leaves that were fermented over a six-week time course. The unfermented samples were characterized by a distinct phyllosphere microbiome, while the endosphere revealed a high similarity. During fermentation, significant microbial shifts were observed, whereby LAB were enhanced in all approaches but never numerically dominated. Among the LAB, Enterococcaceae were identified as the most dominant family in both plants. M. chamomilla community had higher relative abundances of Lactobacillaceae and Carnobacteriaceae, while C. officinalis showed a higher presence of Leuconostocaceae and Streptococcaceae. The natural leaf microbiome and the indigenous LAB communities of field-grown Asteraceae medicinal plants are plant-specific and habitat-specific and are subjected to significant shifts during fermentation. Leaf surfaces as well as leaf endospheres were identified as sources for biopreservative LAB.

Improved antioxidative and cytotoxic activities of chamomile (Matricaria chamomilla) florets fermented by Lactobacillus plantarum KCCM 11613P.

Antioxidative and cytotoxic effects of chamomile (Matricaria chamomilla) fermented by Lactobacillus plantarum were investigated to improve their biofunctional activities. Total polyphenol (TP) content was measured by the Folin-Denis method, and the antioxidant activities were assessed by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method and ß-carotene bleaching method. AGS, HeLa, LoVo, MCF-7, and MRC-5 (normal) cells were used to examine the cytotoxic effects by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay. The TP content of fermented chamomile reduced from 21.75 to 18.76 mg gallic acid equivalent (mg GAE)/g, but the DPPH radical capturing activity of fermented chamomile was found to be 11.1% higher than that of nonfermented chamomile after 72 h of fermentation. Following the ß-carotene bleaching, the antioxidative effect decreased because of a reduction in pH during fermentation. Additionally, chamomile fermented for 72 h showed a cytotoxic effect of about 95% against cancer cells at 12.7 mg solid/ml of broth, but MRC-5 cells were significantly less sensitive against fermented chamomile samples. These results suggest that the fermentation of chamomile could be applied to develop natural antioxidative and anticancer products.

Comparisons of diazotrophic communities in native and agricultural desert ecosystems reveal plants as important drivers in diversity.

Diazotrophs provide the only biological source of fixed atmospheric nitrogen in the biosphere. Although they are the key player for plant-available nitrogen, less is known about their diversity and potential importance in arid ecosystems. We investigated the nitrogenase gene diversity in native and agricultural desert soil as well as within root-associated microbiota of medicinal plants grown in Egypt through the combination of nifH-specific qPCR, fingerprints, amplicon pyrosequencing and fluorescence in situ hybridization-confocal laser scanning microscopy. Although the diazotrophic microbiota were characterized by generally high abundances and diversity, statistically significant differences were found between both soils, the different microhabitats, and between the investigated plants (Matricaria chamomilla L., Calendula officinalis L. and Solanum distichum Schumach. and Thonn.). We observed a considerable community shift from desert to agriculturally used soil that demonstrated a higher abundance and diversity in the agro-ecosystem. The endorhiza was characterized by lower abundances and only a subset of species when compared to the rhizosphere. While the microbiomes of the Asteraceae were similar and dominated by potential root-nodulating rhizobia acquired primarily from soil, the perennial S. distichum generally formed associations with free-living nitrogen fixers. These results underline the importance of diazotrophs in desert ecosystems and additionally identify plants as important drivers in functional gene pool diversity.

Contaminação fúngica de plantas medicinais utilizadas em chás / Fungal contamination of medicinal plants used in teas

Este trabalho teve como objetivo avaliar a qualidade microbiológica quanto à presença de fungos filamentosos em ervas do tipo Peumus boldus Molina (Boldo), Pimpinella anisum L. (erva-doce) e Matricaria chamomilla L. (Camomila) comercializadas na cidade de Campina Grande (Paraíba). Nove amostras de cada planta foram analisadas, totalizando 27 amostras, obtidas aleatoriamente nas feiras livres e barracas informais da cidade. Cada 10 g de amostra foram suspensas em 90 mL de solução salina 0,89% estéril, obtendo-se o concentrado, a partir do qual foram realizadas diluições sucessivas. A seguir, uma alíquota de 0,1mL de cada diluição foi semeada em placas de Petri contendo Ágar Sabouraud Dextrose, as quais foram incubadas à temperatura ambiente por cinco a dez dias. Após este período foram contadas as colônias fúngicas presentes e realizados seus respectivos microcultivos para identificação através de microscópio óptico. Foram encontrados fungos toxigênicos como Aspergillus sp., Penicillium sp. e Fusarium sp., estando o primeiro presente em todas as amostras avaliadas. Também foram identificados os fungos Exophiala sp. E Fonsecaea sp., que possuem importância clínica, podendo causar micose. A forma como estas plantas são armazenadas para comercialização e a falta de fiscalização torna questionável a qualidade das plantas comercializadas para obtenção de chás, o que pode comprometer a saúde do consumidor ao utilizar esses produtos na forma de Chá.

The aim of this study was to assess the microbiological quality, with regard to the presence of filamentous fungi, of the tea herbs Peumus boldus Molina (boldo leaf), Pimpinella anisum L. (anise) and Matricaria chamomilla L. (wild chamomile) marketed in the city of Campina Grande (Paraiba State, Brazil). Nine samples of each plant, totaling 27 samples, were randomly collected in street markets and informal stalls in the city. In the laboratory, 10 g of each sample was ground and suspended in 90 mL of 0.89% sterile saline solution, affording the concentrate from which serial dilutions were obtained. Aliquots of 0.1 mL of each dilution were spread on plates of Sabouraud Dextrose Agar, which were then incubated at room temperature for five to ten days. After this period, the fungal colonies were counted, their morphology was analyzed and subcultures were made on slides for identification by optical microscope. Toxigenic fungi such as Aspergillus sp., Penicillium sp. and Fusarium sp. were found, the first being present in all plant samples. The fungi Exophiala sp. and Fonsecaea sp., which are clinically significant as they can cause mycoses, were also identified. The method of storing these plants before sale and the lack of inspection raise questions about the quality of the marketed herbs, suggesting that the health of the consumer who uses these products to make tea could be harmed.

Avaliação farmacognóstica e microbiológica da droga vegetal camomila (Chamomilla recutita L. ) comercializada como alimento em Cascavel - Paraná / Pharmacognostic and microbiological evaluation of the medicinal plant chamomile (Chamomilla recutita L. ) commercialized as food in Cascavel Municipality, Paraná State, Brazil

As plantas medicinais, dentre elas a camomila, têm sido muito utilizadas na forma de droga vegetal pela população em geral, como importantes alternativas alimentícias e terapêuticas. Considerando que a fiscalização sanitária destes produtos é precária, este quadro torna-se preocupante, visto que, um produto em condições inadequadas para consumo pode acarretar vários riscos ao consumidor. Diante disso, este trabalho teve por objetivo realizar avaliação farmacognóstica e microbiológica em quinze amostras de chá de camomila comercializadas na cidade de Cascavel, Paraná. A metodologia adotada para a realização dos testes foi a preconizada pela Farmacopéia Brasileira (1988, 1996, 1998). O estudo de autenticidade revelou que todas as amostras eram constituídas por Chamomilla recutita L., porém na maioria das amostras os capítulos florais apresentavam-se excessivamente destruídos. Seis amostras apresentaram teor de materiais estranhos acima de 5 por cento. Quanto ao doseamento de óleo essencial, todas as amostras foram insatisfatórias, visto que, apresentaram apenas traços de óleos essenciais, ou seja, valores abaixo de 0,4 por cento, sendo este valor o mínimo exigido pela Farmacopéia. Com relação às análises de bolores e leveduras foi verificado que quatro amostras apresentaram valores acima do limite de 10(4) UFC mL-1, conforme preconizado pela Organização Mundial de Saúde. A partir dos dados obtidos concluiu-se que a camomila comercializada na cidade de Cascavel - Paraná apresenta problemas com relação à qualidade, sendo necessário definir medidas adequadas de controle higiênico sanitário ao longo da cadeia de produção, para garantir a qualidade e segurança destes produtos.

Medicinal plants, including chamomile, have been largely used by the general population as important food and therapeutic alternatives. Considering that the sanitary control of such products is precarious, this situation becomes worrisome since a product presenting unsuitable conditions for consumption can lead to several risks to the consumer. Thus, the present study aimed to perform a pharmacognostic and microbiological evaluation of fifteen samples of chamomile tea commercialized in Cascavel Municipality, Paraná State, Brazil. The adopted methodology for the tests was that recommended by the Brazilian Pharmacopoeia (1988, 1996, 1998). The authenticity study indicated that all samples were constituted of Chamomilla recutita L.; however, most samples had excessively destroyed flower capitula. Six samples had levels of strange materials above 5 percent. As regards essential oil evaluation, all samples were unsatisfactory, since they only had essential oil traces, i.e. values under 0.4 percent, which is the lowest value accepted by the Pharmacopoeia. As regards mold and yeast analyses, four samples had values above the limit of 10(4) UFC mL-1, according to that recommended by the World Health Organization. The obtained data suggest that the chamomile commercialized in Cascavel Municipality has problems regarding quality; thus, establishing suitable procedures for sanitary hygienic control in its production chain is needed to assure the quality and the safety of such products.

Presence of Clostridium botulinum spores in Matricaria chamomilla (chamomile) and its relationship with infant botulism.

Nowadays, infant botulism is the most important form of human botulism in some countries. This illness affects infants younger than 52 weeks of age. The infection occurs in the intestinal tract; therefore, ingestion of Clostridium botulinum spores with food is proposed. In some countries, people use chamomile tea as a household remedy for intestinal colics and given this tea to infants. Chamomile can be contaminated with C. botulinum and could be a vehicle of its spores. Our aim was to study the prevalence and spore-load of C. botulinum in chamomile. We analysed 200 samples; the 7.5% of them were contaminated with botulinum spores. However, prevalence of these spores was significantly higher in chamomile sold by weight in herbal stores (unwrapped chamomile) than prevalence in chamomile sold in tea bags (p=0.0055). The spore-load detected in all positive samples was 0.3-0.4 spores per gram of chamomile. We identified C. botulinum types A, B, and F in the 53.3%, 6.7%, and 13.3%, respectively. Chamomile (principally, unwrapped chamomile) is a potencial vehicle of C. botulinum spores, and ingestion of chamomile tea could represent a risk for infant botulism.