Can Early Life Exposure to Permethrin lead to intergenerational effects?

Pesticides are largely used in agriculture against pests and consequently are present in fruits and vegetables. The wide presence of pesticide residues in breast milk underline the risk for the population, focalizing the long-term consequence of early life pyrethroid exposure. The significant presence of pyrethroid metabolites in the urine of population over the world confirms that their presence in food is a global problem. It has been demonstrated that there is a correlation between the environmental exposure to pesticides and the development of neurodegenerative diseases. Neonatal exposure to Permethrin (PERM), a member of the family of synthetic pyrethroids, can induce neurodegeneration (i.e. Parkinson’s –like disease) and it can cause some alterations in striatum of rats, involving both genetic and epigenetic pathways. The aim of this study was to evaluate if the rat offspring (F1 generation) exposed to a low dose of PERM from postnatal day 6 to 21, presents alterations in Nurr1 gene expression as previously observed in early life permethrin treated male rats. Moreover, global DNA methylation was analyzed in untreated early life exposed mothers and offspring (F1 generation). Methods Through Nurr1gene expression analysis and global DNA methylation assessment in both PERM-treated parents and their untreated offspring, we investigated on the prospective intergenerational effect of this pesticide. Results 33% of progeny presents the same Nurr1 alteration as rats exposed to permethrin in early life. A decrease in global genome-wide DNA methylation was measured in mothers exposed in early life to permethrin as well as in their offspring, whereas untreated rats have a hypermethylated genomic DNA. Conclutions Intergenerational PERM-induced damage on progenies has been identified for the first time. On the light of these results, pesticide residues in the food could represent a risk factor for the health of children especially in early life when the brain is still in the developing phase. Further studies are needed to elucidate the molecular mechanisms associated with the damage

Microbiological investigation of Raphanus sativus L. grown hydroponically in nutrient solutions contaminated with spoilage and pathogenic bacteria

The survival of eight undesired (spoilage/pathogenic) food related bacteria (Citrobacter freundii PSS60, Enterobacter spp. PSS11, Escherichia coli PSS2, Klebsiella oxytoca PSS82, Serratia grimesii PSS72, Pseudomonas putida PSS21, Stenotrophomonas maltophilia PSS52 and Listeria monocytogenes ATCC 19114T) was investigated in mineral nutrient solution (MNS) during the crop cycle of radishes (Raphanus sativus L.) cultivated in hydroponics in a greenhouse. MNSs were microbiologically analyzed weekly by plate count. The evolution of the pure cultures was also evaluated in sterile MNS in test tubes. The inoculated trials contained an initial total mesophilic count (TMC) ranging between 6.69 and 7.78 Log CFU/mL, while non-sterile and sterile control trials showed levels of 4.39 and 0.97 Log CFU/mL, respectively. In general, all inoculated trials showed similar levels of TMC in MNS during the experimentation, even though the levels of the inoculated bacteria decreased. The presence of the inoculums was ascertained by randomly amplified polymorphic DNA (RAPD) analysis applied on the isolates collected at 7-day intervals. At harvest, MNSs were also analyzed by denaturing gradient gel electrophoresis (DGGE). The last analysis, except P. putida PSS21 in the corresponding trial, did not detect the other bacteria, but confirmed that pseudomonads were present in un-inoculated MNSs. Despite the high counts detected (6.44 and 7.24 CFU/g), only C. freundii PSS60, Enterobacter spp. PSS11 and K. oxytoca PSS82 were detected in radishes in a living form, suggesting their internalization

Effect of the natural winemaking process applied at industrial level on the microbiological and chemical characteristics of wine.

The composition of yeast and lactic acid bacteria (LAB) communities and the chemical evolution of the large-scale commercial vinification of Catarratto IGT Sicilia, carried out under the biological regime, was followed from grape harvest until bottling. Simultaneously to the maximumgrowth of yeasts, LAB counts reached high level of concentration (6e7 log CFU mLL1) during the first steps of the alcoholic fermentation. Yeast identification was determined applying different molecular methods. The highest species biodiversity was observed on grape and must samples taken soon after pressing. Saccharomyces cerevisiae was detected at dominant concentrations during the entire winemaking process. LAB cultures were grouped and identified by a combined phenotypic and genotypic approach. Leuconostoc mesenteroides, Lactobacillus hilgardii and Lactobacillus plantarum species were identified; the last was the main LAB recognized during vinification. The winemaking process was also chemically monitored. The alcoholic content was approximately 12.67% (v vL1) at bottling; pH, volatile acidity and total acidity showed a moderate increase during vinification. Tartaric, citric and malic acids decreased until bottling, while lactic acid showed a rapid increase at the end of maceration and bottling. Trans-caffeil tartaric acid was the most abundant phenolic compound and volatile organic compounds (VOC) were mainly represented by isoamylic alcohol, isobutanol, ethyl acetate and octanoic acid

In vivo application and dynamics of lactic acid bacteria for the four-season production of Vastedda-like cheese.

Twelve lactic acid bacteria (LAB), previously selected in vitro (Gaglio et al., 2014), were evaluated in situ for their potential to act as starter cultures for the continuous four-season production of Vastedda-like cheese, madewith raw ewes' milk. The strains belonged to Lactobacillus delbrueckii, Lactococcus lactis subsp. cremoris, Leuconostoc mesenteroides subsp. mesenteroides and Streptococcus thermophilus. LABwere first inoculated in multiple-strain combinations on the basis of their optimal growth temperatures in three process conditions which differed for milk treatment and medium for strain development: process 1, growth of strains in the optimal synthetic media and pasteurised milk; process 2, growth of strains in whey based medium (WBM) and pasteurised milk; and process 3, growth of strains in WBM and raw milk. The strains that acidified the curds in short time, as shown by a pH drop, were all mesophilic and were then tested in a single inoculum through process 3. Randomly amplified polymorphic DNA (RAPD)-PCR analysis applied to the colonies isolated from the highest dilutions of samples confirmed the dominance of the added strains after curd acidification, stretching and storage. After 15 days of refrigerated storage, the decrease in pH values showed an activity of the mesophilic strains at low temperatures, but only Lc. lactis subsp. cremoris PON153, Ln. mesenteroides subsp. mesenteroides PON259 and PON559 increased their number during the 15 days at 7 \ub0C. A sensory evaluation indicated that the cheeses obtained by applying protocol 3 and by inoculation with lactococci are the most similar to the protected denomination of origin (PDO) cheese and received the best scores by the judges. Thus, the experimental cheeses obtained with raw milk and inoculated with single and multiple combinations of lactococci were subjected to the analysis of the volatile organic compounds (VOCs) carried out by a headspace solid phase microextraction (SPME) technique coupled with gas chromatography with mass spectrometric detection (GC/MS). The dominance of lactococci over thermophilic LAB of raw milk was verified during summer production and, based on the combination of VOC profiles and sensory evaluation of the final cheeses, the multi-strain Lactococcus culture resulted in the most suitable starter preparation for the full-year production of Vastedda-like cheese

Identification, typing, and investigation of the dairy characteristics of lactic acid bacteria isolated from 'Vastedda della valle del Belìce' cheese

Traditional cheeses made without starter cultures can be characterised by the attribute of instability. The addition of autochthonous starter cultures can ensure stability without compromising the characteristics of the final product. This study aimed to characterise the autochthonous lactic acid bacteria (LAB) population in “Vastedda della valle del Belìce” cheeses, which have a protected designation of origin (PDO) status, in order to develop an ad hoc starter culture to be used in its future production. Winter and spring productions were analysed to ensure isolation of specific LAB that had adapted to perform fermentation at low temperatures. Plate counts revealed total microbial numbers nearing 109 CFU.g−1. All of the cheese samples were dominated by coccus-shaped LAB. When enterobacteria were present, their concentrations were at similar levels (3.3–5.6 Log CFU.g−1) in both seasons. All of the colonies that differed in morphological appearance were isolated and differentiated on the basis of phenotypic characteristics and genetic polymorphisms, as analysed by random amplification of polymorphic DNA-polymerase chain reaction. A total of 74 strains were identified and further genotyped by sequencing the 16S rRNA gene, resulting in the identification of 16 LAB species belonging to five genera (Enterococcus, Lactobacillus, Lactococcus, Leuconostoc and Streptococcus). The species most frequently present were Streptococcus gallolyticus subsp. macedonicus, Streptococcus thermophilus, Lactococcus lactis and Leuconostoc mesenteroides. The 74 strains were also investigated in vitro for general dairy parameters such as acidification capacity, diacetyl generation and antibacterial activity. Several strains of the most frequently represented species displayed traits relevant to the production of PDO “Vastedda della valle del Belìce”

Animal rennets as sources of dairy lactic acid bacteria

The microbial composition of artisan and industrial animal rennet pastes was studied by using both culture-dependent and -independent approaches. Pyrosequencing targeting the 16S rRNA gene allowed to identify 361 operational taxonomic units (OTUs) to the genus/species level. Among lactic acid bacteria (LAB), Streptococcus thermophilus and some lactobacilli, mainly Lactobacillus crispatus and Lactobacillus reuteri, were the most abundant species, with differences among the samples. Twelve groups of microorganisms were targeted by viable plate counts revealing a dominance of mesophilic cocci. All rennets were able to acidify ultrahigh-temperature-processed (UHT) milk as shown by pH and total titratable acidity (TTA). Presumptive LAB isolated at the highest dilutions of acidified milks were phenotypically characterized, grouped, differentiated at the strain level by randomly amplified polymorphic DNA (RAPD)-PCR analysis, and subjected to 16S rRNA gene sequencing. Only 18 strains were clearly identified at the species level, as Enterococcus casseliflavus, Enterococcus faecium, Enterococcus faecalis, Enterococcus lactis, Lactobacillus delbrueckii, and Streptococcus thermophilus, while the other strains, all belonging to the genus Enterococcus, could not be allotted into any previously described species. The phylogenetic analysis showed that these strains might represent different unknown species. All strains were evaluated for their dairy technological performances. All isolates produced diacetyl, and 10 of them produced a rapid pH drop in milk, but only 3 isolates were also autolytic. This work showed that animal rennet pastes can be sources of LAB, mainly enterococci, that might contribute to the microbial diversity associated with dairy productions

Molecular signatures associated with the treatment of triple-negative MDA-MB231 breast cancer cells with the histone deacetylase inhibitors JAHA and SAHA

Jay Amin Hydroxamic Acid (JAHA; N8-ferrocenylN1-hydroxy-octanediamide) is a ferrocene-containing analogue of the histone deacetylase inhibitor (HDACi) suberoylanilide hydroxamic acid (SAHA). JAHA’s cytotoxic activity on MDA-MB231 triple negative breast cancer (TNBC) cells at 72 h has been previously demonstrated with an IC50 of 8.45 M. JAHA’s lethal effect was found linked to perturbations of cell cycle, mitochondrial activity, signal transduction and autophagy mechanisms. In order to glean novel insights on how MDA-MB231 breast cancer cells respond to the cytotoxic effect induced by JAHA, and to compare the biological effect with the related compound SAHA, we have employed a combination of differential display-PCR, proteome analysis and COMET assay techniques and shown some differences in the molecular signature profiles induced by exposure to either HDACis. In particular, in contrast to the more numerous and diversified changes induced by SAHA, JAHA has shown a more selective impact on expression of molecular signatures involved in anti-oxidant activity and DNA repair. Besides expanding the biological knowledge of the effect exerted by the modifications in compound structures on cell phenotype, the molecular elements put in evidence in our study may provide promising targets for therapeutic interventions on TNBCs

Ecology of Vastedda della valle del Belìce cheeses: A review and recent findings to stabilize the traditional production

The increasing request for classic foods has led to a higher demand for cheeses enjoying a “recognition of quality” status. This phenomenon has also affected Sicilian dairy products, in particular protected designation of origin (PDO) “Vastedda della valle del Belìce” (VdB), a cheese traditionally available only during the summer season, but requested year round. The variations of the microbial populations of raw milk among the seasons influence the microbiological quality of the cheeses produced. This review article reports the historical importance of cheeses and the traditional equipment used for dairy productions in Sicily with a specific focus on VdB. The microbial ecology of this cheese is thoroughly discussed. The recent findings to stabilize the traditional VdB production protocol with regards to the microbial composition and the organoleptic characteristics of the final cheeses are reviewed. The selection of starter lactic acid bacteria from PDO cheeses, their in vitro and in vivo application, and, overall, the setup of their inoculation, have all generated a novel production protocol. The innovation was respectful of the traditional discipline, essentially based on the ad hoc biofilm formation on the traditional wooden equipment, and allowed the preservation of VdB typicality. Future prospects are summarized