Antiproliferative, Proapoptotic, Antioxidant and Antimicrobial Effects of Sinapis nigra L. and Sinapis alba L. Extracts.

High Brassicaceae consumption reduces the risk of developing several cancer types, probably due to high levels of glucosinolates. Extracts from Sinapis nigra L. (S. nigra) and Sinapis alba L. (S. alba) have been obtained from leaves and seeds under different conditions using ethanol/water mixtures because their glucosinolates are well accepted by the food industry. The EtOH/H2O 8:2 mixture gives better yields in glucosinolate amounts from ground seeds, mainly, sinalbin in S. alba and sinigrin in S. nigra. The highest antiproliferative activity in both non-tumor and tumor cell lines was induced by S. alba seeds extract. To evaluate whether the effect of Sinapis species (spp) was only due to glucosinolate content or whether it was influenced by the extracts' complexity, cells were treated with extracts or glucosinolates, in the presence of myrosinase. Pure sinigrin did not modify cell proliferation, while pure sinalbin was less effective than the extract. The addition of myrosinase increased the antiproliferative effects of the S. nigra extract and sinigrin. Antiproliferative activity was correlated to Mitogen-Activated Protein Kinases modulation, which was cell and extract-dependent. Cell-cycle analysis evidenced a proapoptotic effect of S. alba on both tumor cell lines and of S. nigra only on HCT 116. Both extracts showed good antimicrobial activity in disc diffusion tests and on ready-to-eat fresh salad. These results underline the potential effects of Sinapis spp in chemoprevention and food preservation.

Aerosol exposure and risk assessment for green jobs involved in biomethanization.

Anaerobic digestion is a consolidated biotechnology able to produce renewable energy from biomasses. In the European countries, quick growth of biogas production from different organic matrices including wastes has been observed. In relation to the characteristics and quantity of the anaerobic digestion of feedstock, there are different technologies, advantages and criticisms. An accurate occupational risk assessment and development of management tools for green jobs involved in the anaerobic digestion plants are due. The aim of this work is to assess the aerosol exposure for such workers, focusing on the bioaerosol risk. Full scale plants for the treatment of organic municipal waste, waste water treatment sludge, agro zootechnical and food producing byproducts were involved for this purpose. The bioaerosol levels were monitored during activities through culturing and biomolecular methods; moreover, the sub-fractionated PM10 and carried endotoxins were measured in different plant areas. Global microbial contamination is higher (>5000 UFC/m3) in the area where organic wastes are handled and pretreated, both for organic municipal waste plants - with a bacterial prevalence - and agro zootechnical plants - with a fungi prevalence. Moreover, the microbial contamination is higher where organic municipal waste is present in respect to other biomasses (ANOVA p < 0.01). Numerous pathogens are carried by the aerosol. HAdV-4 presence is lower than LOQ (50 gene copies/m3) in all the samples. Environmental PM10 reached the 280 µg/m3 level including PM3 for 78%. Endotoxin pollution overtakes the 90 EU/m3 limit sporadically. Personal PM4.5 reached 10 mg/m3 only for maintenance technicians in the pretreatment area for organic municipal waste. The risk can be evaluated under a quantitative and qualitative point of view highlighting risk management improvement for anaerobic digestion plants.

Assessing Methanobrevibacter smithii and Clostridium difficile as not conventional faecal indicators in effluents of a wastewater treatment plant integrated with sludge anaerobic digestion.

Wastewater treatment plants (WWTP) are an important source of surface water contamination by enteric pathogens, affecting the role of environmental water as a microbial reservoir. We describe the release to the environment of certain anaerobes of human and environmental concern. The work was focused on emerging microbial targets. They are tracing, by RT-qPCR, on WWTP effluents, both liquid and solid, when an anaerobic digestion step is included. The focus is placed on Clostridium spp. with the specific quantification of Clostridium perfringens, as typical bioindicator, and Clostridium difficile, as emerging pathogen not only confined into nosocomial infection. Moreover methanogens were quantified for their involvement in the anaerobic digestion, and in particular on Methanobrevibacter smithii as major methanogenic component of the human gut microbiome and as not conventional faecal indicator. In the water samples, a reduction, statistically significant, in all microbial targets was observed (p < 0.01), 2 log for the total bacteria, 1.4 log for the Clostridium spp. and M. smithii, 1 log for total methanogens, C. perfringens and C. difficile. The AD process contribute to a significant change in microbial levels into the sludge for total bacteria and total methanogens (p < 0.01), both when the input sludge are primary and secondary, while for the presence of Clostridium spp. and C. difficile there was not a significant change. The produced data are innovative showing which is the diffusion of such anaerobic microorganisms throughout the WWTP and opening a discussion on the implementation of possible techniques for a more efficient microbial removal from effluents, particularly bio-solids, to reduce the potential release of pathogens into the environment.

Probiotics and gut health in infants: A preliminary case-control observational study about early treatment with Lactobacillus reuteri DSM 17938.

AIMS: We performed this case-control observational study to evaluate the effects of early administration of Lactobacillus reuteri DSM 17938 on microbial composition in infants' gastrointestinal tract. METHODS: Early fecal microbiota composition was analyzed by using selective and differential cultural methods. Genomic DNA from positive Escherichia coli and Cronobacter sakazakii colonies was extracted and DNA was processed by multiplex PCR assay. RESULTS: Fecal samples of 30 hospitalized infants who previously received probiotics and 30 not receiving probiotics were analyzed. We find that the two groups showed differences in gut microbial strains composition and richness. Infant treated with probiotics have a lower total anaerobic gram negative counts (p=0.03) and a higher total anaerobic gram-positive counts (p=0.02). Enterobacteriaceae and enterococci were significantly higher (p=0.04) in the control group. No significant differences were observed for total aerobic counts, lactobacilli and bifidobacteria. C. sakazaki was found only in one infant recruited in the control group. Infants not previously treated with probiotics showed a higher colonization by diarrheagenic E. coli (EPEC) (p=0.04). CONCLUSIONS: Our findings enhanced our understanding of the effects of probiotics on gut health in pediatric subjects. Early administration of L. reuteri in infancy could improve gut health by reducing pathogens colonization.

Diarrheagenic Escherichia coli in acute gastroenteritis in infants in North-West Italy.

UNLABELLED: Enteropathogenic Escherichia coli may cause diarrhoea in infancy, but it is not routinely detected and regarded as a major causative agent. The aim of the present study was to estimate the incidence of enteropathogenic E. coli infection and to investigate its epidemiology and pathogenesis from faecal specimens in infants hospitalized for acute gastroenteritis. Between March 2008 and June 2009, faecal samples were collected and examined to recognize diarrhoeal aetiology, especially for E. coli, by cultural identification and multiplex-PCR. E. coli were isolated in 75 of 160 collected samples (46,88%); 10 samples of which (6,3%) had been positively recognised for pathogenic genes. Data showed that the presence of diarrheagenic E. coli infection was 6.3%, but it becomes 5% considering E. coli as a unique agent responsible for diarrhoea. The datum is not statistically meaningful because of the small sample (p>0,05). Bacterial pathogens were also isolated in 60 samples (37,5% of the total collected samples): 15 Salmonella spp., 8 Klebsiella pneumoniae, 9 Klebsiella oxytoca, 11 Citrobacter freundii, 5 Pseudomonas aeruginosa, 2 Serratia spp., 7 Enterobacter cloacae, 1 Shigella spp., 2 Campylobacter spp. Rotavirus was the predominant pathogenic single etiologic agent identified. It was found in 35 samples (21.88% of the overall collected samples), while Adenovirus, serotypes 40 or 41, was isolated in 2 samples (1.3%). Rotavirus infection was found predominantly in winter with respect to autumn. Data provide an interesting epidemiologic survey of enteropathogenic E. coli, which is not usually detected, although it may have potential clinical implications. ABBREVIATIONS: CDEC, detaching E. coli; DAEC, diffusely adherent E. coli; EAggEC, enteroaggregative E. coli; EHEC, enterohaemorrhagic E. coli; EIEC, enteroinvasive E. coli; EPEC, enteropathogenic E. coli; ETEC, enterotoxigenic E. coli.