Technological characterization of bacteriocin producing Lactococcus lactis strains employed to control Listeria monocytogenes in cottage cheese

In recent years, there has been a particular focus on the application of antimicrobial compounds produced by lactic acid bacteria (LAB) as natural preservatives to control the growth of spoilage and pathogenic bacteria in food. Bacteriocins are antimicrobial peptides which can be added to foods in concentrated forms as food preservatives, e.g. additives, or they can be produced in situ by starters or protective cultures. In this study, twenty Lactococcus lactis bacteriocin producers previously isolated from Italian fermented foods were subjected to a variety of physical and biochemical tests in order to identify those with the greatest potential as starter cultures in cheese production. Of these, four strains isolated from cheese (one nisin Z producer, one nisin A producer and two lacticin 481 producers) which fulfilled the desired technological criteria were assessed for their ability to control Listeria monocytogenes. The subsequent application of these bacteriocinogenic strains as starter cultures in Cottage cheese established that the nisin A producing Lact. lactis 40FEL3, and to a lesser extent the lacticin 481 producers 32FL1 and 32FL3, successfully controlled the growth of the pathogen. This is the first study to directly compare the ability of nisin A, nisin Z and lacticin 481 producing strains to control listerial growth during the manufacture and storage of Cottage cheese.     

Temperature and storage effects on antioxidant activity of juice from red and white grapes

The effect of temperature and storage time on the phenolic composition and on the antioxidant activity of grape (Vitis vinifera L.) juices made using a red (Sangiovese, SG) and a white (Muscat of Alexandria, MA) variety was studied. Total phenolics, flavonoids, flavan‐3‐ols and hydroxycinnamoyl‐tartrates (HCT) were determined on grape extracts (GE) and juices. Total anthocyanins and anthocyanins composition were measured on GE. The antioxidant activity was assessed by means of two different in vitro tests: scavenging of the stable 2,2‐diphenyl‐1‐picrylhydrazyl radical and the inhibition of tyrosine nitration mediated by authentic peroxynitrite (ONOO^?). All the juices were analysed after 24 h and 2 weeks of storage by means of two‐way anova (factors being cultivar and temperature). Anthocyanins were not detected in MA extract, whereas in SG their content was 534 mg malvidin‐equivalent per 100 g of dry matter (d.m.) and malvidin derivatives (glycosylated and coumaroylated) were the most represented (respectively 169 and 41 malvidin‐equivalent per 100 g of d.m.). HCT content of the extracts was higher in SG (overall +33%). Also in grape juices, HCT were lower in MA and in this variety, the trans‐fertaric and cis‐coutaric acids were also undetectable. Cultivar effect proved to be highly significant, while no significant differences in the phenolic composition were observed for storage temperatures (4 and ?20 °C) and cultivar × temperature interaction. However, when statistical analysis was focused on each cultivar, MA was found to be more sensible to storage conditions and a significant reduction in total phenolics (?20%) and flavonoids (?53%) content and in the ONOO^? scavenging potential (?32%) was evident after 2 weeks at 4 °C (when compared with the same storage temperature after 24 h). On the contrary, SG juices did not show significant differences among the four storage treatments investigated. These results could be explained suggesting that anthocyanins presence in red grape plays a key role in juice stability.     

Lactic acid bacteria ecology of three traditional fermented sausages produced in the North of Italy as determined by molecular methods

In this study the bacterial biodiversity during the maturation process of three traditional sausages produced in the North of Italy (Salame bergamasco, Salame cremonese and Salame mantovano) was investigated by using culture-dependent and -independent methods. Eleven plants, in the three provinces considered here, were selected because starter cultures were never used in the production. The bacterial ecology, as determined by plate counts, was dominated by lactic acid bacteria (LAB), with minor contribution of coagulase negative cocci and yeasts. After molecular identification of 486 LAB strains, the species more frequently isolated were Lactobacillus sakei and Lactobacillus curvatus. This evidence was also confirmed by PCR-Denaturing Gradient Gel Electrophoresis (DGGE). All the samples analyzed were characterized by the constant presence of L. sakei and L. curvatus bands. A richer biodiversity was only detected at the beginning of maturation. The results obtained by the molecular characterization of the L. sakei and L. curvatus and by the cluster analysis of the DGGE profiles highlighted a plant-specific population, rather than a geographic characterization of the products, underlining how the environmental and processing conditions are able to select specific microbiota responsible for the main transformations during the fermentation and ripening of the sausages.     

Identification and analysis of isothiocyanates and new acylated anthocyanins in the juice of Raphanus sativus cv. Sango sprouts

The freeze-dried sprouts’ juice of Raphanus sativus (L.) cv. Sango was prepared and analysed for the first time. HPLC analysis of total isothiocyanates, after protein displacement, resulted in 77.8 ± 3.0 μmol/g of dry juice while GC–MS analysis of hexane and acetone extracts showed E- and Z-raphasatin (8.9 and 0.11 μmol/g, respectively) and sulforaphene (11.7 μmol/g), summing up to 20.7 ± 1.7 μmol/g of free isothiocyanates. Sprouts’ juice contained an unprecedented wealth of anthocyanins and a new fractionation methodology allowed us to isolate 34 mg/g of acylated anthocyanins (28.3 ± 1.9 μmol/g), belonging selectively to the cyanidin family. Analysis was performed by HPLC–PDA–ESI–MSⁿ and extended to deacylated anthocyanins and aglycones, obtained, respectively, by alkaline and acid hydrolysis. This study identified 70 anthocyanins, 19 of which have never been described before and 32 of which are reported here in R. sativus for the first time. Sango radish sprouts are exceptional dietary sources of heath-promoting micronutrients.     

用于地毯和纺织品的新纤维

在意大利Aquafil公司与特伦托大学合作进行的科研工作的基础上,介绍和讨论了用于地毯和纺织品的新纤维的新的试验方法.此外,指出了各种创新的多种聚合物复合所得的结果.结果证实,由反应纺丝得到的多种聚合物的复合纤维,力学性质具有单值性.     

Online Portable Microcantilever Biosensors for Salmonella enterica Serotype Enteritidis Detection

The micro- and nano-technologies coupled with a deep knowledge of organic/inorganic interfaces guarantee an exceptional sensitivity and specificity of the sensor, while the lab-on-a-chip platform reduces assay times and limits sampling and/or sample preparation, providing compact and portable objects. Therefore, the development of innovative biosensors such as antibody-immobilized microcantilevers can overcome the evident limits of nowadays technologies, such as time consuming, expensiveness, difficult automation, low sensitivity, accuracy, and precision for quantitative methods. The present study proposes two device designs for the detection of food pathogens, exploiting an antibody-immobilized microcantilever biosensors, a novel class of mass detectors. For the first one, we integrated the mechanical sensors on a microfluidic platform (lab-on-a-chip) to perform online analysis, directly in liquid environment. We showed that our portable biosensors could easily detect the presence of pathogenic bacteria such as Salmonella enterica serotype enteritidis in concentration 10⁵ cfu/mL in just 40 min, without any enrichment and/or sample preparation. To increase the mass sensitivity of our analysis, we also fabricated microstructures optimized for vibrating in vacuum environment. Using a dip-and-dry technique, we showed that, in such configuration, the experimental limit of detection is as low as 10³ cfu/mL. Due to the extremely small volumes needed, our biosensors operating in vacuum have the potentiality of detecting the presence or absence of a single cell.     

Isolation and characterization of a psychrotolerant toxin producer, Bacillus weihenstephanensis, in liquid egg products

A psychrotolerant bacteria of the Bacillus cereus group was found responsible for the spoilage of whole liquid egg products. By sequencing a 16S rRNA region and performing a PCR amplification of specific 16S rRNA and cspA signatures, a Bacillus weihenstephanensis was identified. Characterization of this strain shows its ability to grow in defined medium as well as in whole liquid egg at refrigerated temperatures. The strain isolated possesses genes encoding for hemolysin BL, nonhemolytic enterotoxin, and B. cereus enterotoxins and produces enterotoxins with cytotoxic activity in whole liquid egg, even at refrigerated temperatures. The isolate exhibits a clear ability to stick and form biofilms on stainless steel, the most common material used in egg breaking factories, as well as on model hydrophilic (glass) and hydrophobic (polytetrafluoroethylene) materials. These findings show the necessity to monitor for Bacillus contamination in egg products that are often used in the composition of particularly susceptible finished products such as cream, dessert, dairy, meat, and seafood.     

Detection of glyphosate and its metabolites in food of animal origin based on ion-chromatography-high resolution mass spectrometry (IC-HRMS)

Glyphosate and glufosinate are broad spectrum herbicides, widely used in agriculture and in inhabited or industrialised areas, and aminomethylphosphonic acid is a degradation product of glyphosate. In 2015, the International Agency for Research on Cancer reported that glyphosate is a probable carcinogenic. In 2017, however, a scientific opinion of the European Chemicals Agency concluded that glyphosate is not proven to be carcinogenic, mutagenic or to have negative effects on reproduction. Nevertheless, aminomethylphosphonic acid was not considered. Due to their chemical-physical characteristics, these molecules present difficulties that have not yet allowed routine monitoring to be carried out. For these reasons, we developed and validated a simple and versatile liquid extraction, before IC-HRMS analysis, of three different complex matrices: honey, bass fish and bovine muscle. Among the satisfactory validation parameters, the LOQs in the range of 4.30–9.26 ng g^?1 demonstrated high method sensitivity, compared to the few works present in literature. Finally, the method was applied to real commercial samples, which showed no traces of the selected pesticides.     

Detection, quantification and vitality of Listeria monocytogenes in food as determined by quantitative PCR

In this paper we describe the development of a quantitative PCR (qPCR) technique to detect, quantify and determine the vitality of Listeria monocytogenes in foods. The method was based on the amplification of the intergenic region spacer (IGS) between the 16S and 23S rRNA genes. A panel of more than 100 strains of Listeria spp. and non-Listeria was used in order to verify the specificity of the primers and Taqman probe and amplification signals were obtained only when L. monocytogenes DNA and RNA were loaded in the qPCR mix. Standard curves were constructed in several food matrices (milk, meat, soft cheese, fermented sausage, cured ham and ready-to-eat salad). The quantification limit was of 10(3)-10(4) cfu/g or ml, while for the determination of vitality it was 10(4)-10(5) cfu/g or ml. After an overnight enrichment in BHI at 37 degrees C also 10 cfu/g or ml could be detected in all the matrices used in this study. When we applied the protocol to food samples collected from the market or from small food processing plants, on a total number of 66 samples, 4 fresh cheeses from raw milk gave positive results prior to the overnight incubation, while 9 samples, of which only one represented by fresh meat and the others by cheeses from raw milk, were positive after the enrichment. Out of the 4 positive samples, only one could be quantified and it was determined to contain 4x10(3) cfu/g.     

Spoilage of blood sausages morcilla de Burgos treated with high hydrostatic pressure

In this study, the microbial ecology of the blood sausage morcilla de Burgos, subjected to high hydrostatic pressure treatment (HPP), was studied by culture-dependent and -independent methods. Morcilla de Burgos is the most traditional and famous blood sausage in Spain. The producers are interested in extending its shelf-life in order to expand their market and to reduce losses attributed to spoilage. Sausage batter prior to stuffing and blood sausages HPP treated or not (control) were analyzed at 0, 9, 14, 21, 28 and 35 days of storage at 4 degrees C. Lactic acid bacteria, Pseudomonas spp. and aerobic mesophilic bacteria were investigated by traditional plating. PCR-denaturing gradient gel electrophoresis (DGGE) was used to analyze the DNA and the RNA extracted directly from the blood sausages, as well as bulk cells of LAB and Pseudomonas spp. The results showed that HPP improved the shelf life of morcilla de Burgos to 28 days in comparison with control samples. The populations responsible for spoilage, namely LAB, remained lower in HPP treated samples when compared with the control samples. Only at 35 days of storage they reached values of 10(8) cfu/g, leading to the spoilage of the product. Although, HPP affected the LAB population, they were able to recover the injury provoked by the treatment. Lastly, HPP seemed to affect differently LAB species detected. While Leuconostoc mesenteroides was completely inactivated by HPP, Weissella viridescens was able to recover and carry out the typical spoilage of the product. Pseudomonas spp. remained under detection level (<10(2) CFU/g) after the HPP treatment.