Effectiveness of lactic and acetic acids on the growth of Listeria monocytogenes and Bacillus cereus in primo sale fresh cheese

In this study Brain Heart Infusion broth inoculated with Listeria monocytogenes with a concentration of acetic acid of 24.98 mM or a concentration of 44.40 mM of lactic acid did not determine the increase in absorbance in 7 days. A concentration of acetic acid of 24.98 mM and a concentration of 22.20 mM of lactic acid were effective against Bacillus cereus growth. Then, challenge tests on primo sale cheese were conducted to establish if these concentrations were efficient when applied to cheese. After inoculum with the pathogens (2 log CFU g−1), cheese was dipped with acetic and lactic acid solutions. In a first trial, L. monocytogenes inoculated, showed the absence of significant differences in growth at 4 °C among the treated series (Ac1: acetic acid - 49.96 mM; Ac2: 24.98 mM; Lac1: lactic acid – 88.80 mM; Lac2: 44.40 mM) if compared to Control (CTRL) series (dipped with sterile water). At 8 °C, a significantly lower growth in Ac1 samples if compared to control ones and to all the treated series was observed (P < 0.05). The trial conducted inoculating B. cereus did not show any difference at 15 °C among samples treated with organic acids if compared to control series

FlhF, a signal recognition particle-like GTPase, is involved in the regulation of flagellar arrangement, motility behaviour and protein secretion in Bacillus cereus

Flagellar arrangement is a highly conserved feature within bacterial species. However, only a few genes regulating cell flagellation have been described in polar flagellate bacteria. This report demonstrates that the arrangement of flagella in the peritrichous flagellate Bacillus cereus is controlled by flhF. Disruption of flhF in B. cereus led to a reduction in the number of flagella from 10-12 to 1-3 filaments per cell in the insertion mutant MP06. Moreover, compared to the parental strain, MP06 exhibited: (i) shorter smooth swimming phases, causing reduced swimming motility but not affecting chemotaxis; (ii) complete inhibition of swarming motility, as differentiated swarm cells were never detected; (iii) an increased amount of extracellular proteins; and (iv) differential export of virulence determinants, such as haemolysin BL (HBL), phosphatidylcholine-preferring phospholipase C (PC-PLC) and non-haemolytic enterotoxin (NHE). Introduction of a plasmid harbouring flhF (pDGflhF) into MP06 completely restored the wild-type phenotype in the trans-complemented strain MP07. B. cereus flhF was found to constitute a monocistronic transcriptional unit and its overexpression did not produce abnormal features in the wild-type background. Characterization of a B. cereus mutant (MP05) carrying a partial flhF deletion indicated that the last C-terminal domain of FlhF is involved in protein export while not required for flagellar arrangement and motility behaviour. Taken together, these data suggest that B. cereus FlhF is a promising candidate for connecting diverse cellular functions, such as flagellar arrangement, motility behaviour, pattern of protein secretion and virulence phenotype

Spotlight on the Compositional Quality of Probiotic Formulations Marketed Worldwide

On the worldwide market, a great number of probiotic formulations are available to consumers as drugs, dietary supplements, and functional foods. For exerting their beneficial effects on host health, these preparations should contain a sufficient amount of the indicated living microbes and be pathogen-free to be safe. Therefore, the contained microbial species and their amount until product expiry are required to be accurately reported on the labels. While commercial formulations licensed as drugs are subjected to rigorous quality controls, less stringent regulations are generally applied to preparations categorized as dietary supplements and functional foods. Many reports indicated that the content of several probiotic formulations does not always correspond to the label claims in terms of microbial identification, number of living organisms, and purity, highlighting the requirement for more stringent quality controls by manufacturers. The main focus of this review is to provide an in-depth overview of the microbiological quality of probiotic formulations commercialized worldwide. Many incongruences in the compositional quality of some probiotic formulations available on the worldwide market were highlighted. Even if manufacturers carry at least some of the responsibility for these inconsistencies, studies that analyze probiotic products should be conducted following recommended and up-to-date methodologies

Microbiological quality and resistance to an artificial gut environment of two probiotic formulations

The quality control of probiotic products is the focus of numerous organizations worldwide. Several studies have highlighted the poor microbiological quality of many commercial probiotic formulations in terms of the identity of the contained microorganisms, viability, and purity, thus precluding the expected health benefits and representing a potential health risk for consumers. In this paper, we analyzed the contents of two probiotic formulations, one composed of an encapsulated mixture of lactobacilli and bifidobacteria, and one by a lyophilized yeast. The microorganisms contained in the products were quantified and identified using up-to-date methodologies, such as MALDI-TOF MS and metagenomic analysis. Moreover, as acid and bile tolerance is included among the criteria used to select probiotic microorganisms, in vitro tests were performed to evaluate the behavior of the formulations in conditions mimicking the harsh gastric environment and the intestinal fluids. Our results indicate the high quality of the formulations in terms of the enumeration and identification of the contained organisms, as well as the absence of contaminants. Moreover, both products tolerated the acidic conditions well, with encapsulation providing further protection for the microorganisms. A good tolerance to the simulated artificial intestinal conditions was also evidenced for both preparations

Contribution of Surfactin and SwrA to Flagellin Expression, Swimming, and Surface Motility in Bacillus subtilis.

Multicellular communities produced by Bacillus subtilis can adopt sliding or swarming to translocate over surfaces. While sliding is a flagellum-independent motility produced by the expansive forces in a growing colony, swarming requires flagellar functionality and is characterized by the appearance of hyperflagellated swarm cells that associate in bundles or rafts during movement. Previous work has shown that swarming by undomesticated B. subtilis strains requires swrA, a gene that upregulates the expression of flagellar genes and increases swimming motility, and surfactin, a lipopeptide biosurfactant that also facilitates sliding. Through an analysis of swrA(+) and swrA mutant laboratory strains with or without a mutation in sfp (a gene involved in surfactin production), we show that both swrA and surfactin upregulate the transcription of the flagellin gene and increase bacterial swimming. Surfactin also allows the nonswarming swrA mutant strain to efficiently colonize moist surfaces by sliding. Finally, we reconfirm the essential role of swrA in swarming and show that surfactin, which increases surface wettability, allows swrA(+) strains to produce swarm cells on media at low humidity

Transport layer protocols and architectures for satellite networks

Designing efficient transmission mechanisms for advanced satellite networks is a demanding task, requiring the definition and the implementation of protocols and architectures well suited to this challenging environment. In particular, transport protocols performance over satellite networks is impaired by the characteristics of the satellite radio link, specifically by the long propagation delay and the possible presence of segment losses due to physical channel errors. The level of impact on performance depends upon the link design (type of constellation, link margin, coding and modulation) and operational conditions (link obstructions, terminal mobility, weather conditions, etc.). To address these critical aspects a number of possible solutions have been presented in the literature, ranging from limited modifications of standard protocols (e.g. TCP, transmission control protocol) to completely alternative protocol and network architectures. However, despite the great number of different proposals (or perhaps also because of it), the general framework appears quite fragmented and there is a compelling need of an integration of the research competences and efforts. This is actually the intent of the transport protocols research line within the European SatNEx (Satellite Network of Excellence) project. Stemming from the authors' work on this project, this paper aims to provide the reader with an updated overview of all the possible approaches that can be pursued to overcome the limitations of current transport protocols and architectures, when applied to satellite communications. In the paper the possible solutions are classified in the following categories: optimization of TCP interactions with lower layers, TCP enhancements, performance enhancement proxies (PEP) and delay tolerant networks (DTN). Advantages and disadvantages of the different approaches, as well as their interactions, are investigated and discussed, taking into account performance improvement, complexity, and compliance to the standard semantics. From this analysis, it emerges that DTN architectures could integrate some of the most efficient solutions from the other categories, by inserting them in a new rigorous framework. These innovative architectures therefore may represent a promising solution for solving some of the important problems posed at the transport layer by satellite networks, at least in a medium-to-long-term perspective. Copyright (c) 2006 John Wiley & Sons, Ltd

Delivery Mode Shapes the Composition of the Lower Airways Microbiota in Newborns

Radical alterations in the human microbiota composition are well-known to be associated with many pathological conditions. If these aberrations are established at the time of birth, the risk of developing correlated pathologies throughout life is significantly increased. For this reason, all newborns should begin their lives with a proper microbiota in each body district. The present study aimed at demonstrating a correlation between the mode of delivery and the development of a well-balanced microbiota in the lower airways of newborns. 44 pregnant women were enrolled in this study. Microbiological comparative analysis was carried out on tracheobronchial secretions of babies born through vaginal delivery (VD) or caesarean section (CS). All samples showed the presence of bacterial DNA, regardless of the mode of delivery. No viable cultivable bacteria were isolated from the CS samples. On the contrary, VD allowed colonization of the lower airways by alive cultivable bacteria. The identification of bacterial species revealed that Lactobacillus spp. and Bacteroides vulgatus were the most common microorganisms in the lower airways of vaginally-delivered newborns. Data obtained from quantitative PCRs showed a significantly higher total bacterial load, as well as Firmicutes and Lactobacillus spp. amount, in VD samples than CS ones, while no statistically significant difference was found in Torque Teno Virus (TTV) load between samples. Taken together, our findings confirm the hypothesis that passage through the maternal vaginal canal determines more beneficial colonization of the lower airways in newborns

Characterization of a Bacillus cereus strain associated with a large feed-related outbreak of severe infection in pigs

Aims: Bacillus cereus is often responsible for foodborne diseases and both local and systemic infections in humans. Cases of infection in other mammals are rather rare. In this study, we report a B. cereus feed-related outbreak that caused the death of 6234 pigs in Italy. Methods and Results: Massive doses of a Gram-positive, spore-forming bacterium were recovered from the animal feed, faeces of survived pigs and intestinal content of dead ones. The B. cereus MM1 strain was identified by MALDI-TOF MS and typified by RAPD-PCR. The isolate was tested for the production of PC-PLC, proteases, hemolysins and biofilm, for motility, as well as for the presence of genes encoding tissue-degrading enzymes and toxins. Antimicrobial resistance and pathogenicity in Galleria mellonella larvae were also investigated. Our results show that the isolated B. cereus strain is swimming-proficient, produces PC-PLC, proteases, hemolysins, biofilm and carries many virulence genes. The strain shows high pathogenicity in G. mellonella larvae. Conclusions: The isolated B. cereus strain demonstrates an aggressive profile of pathogenicity and virulence, being able to produce a wide range of determinants potentially hazardous to pigs' health. Significance and Impact of Study: This study highlights the proficiency of B. cereus to behave as a devastating pathogen in swine if ingested at high doses and underlines that more stringent quality controls are needed for livestock feeds and supplements