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.

Computational Analysis of Turbulent Hemodynamics in Radiocephalic Arteriovenous Fistulas to Determine the Best Anastomotic Angles.

BACKGROUND: Hemodynamics has been known to play a major role in the development of intimal hyperplasia leading to arteriovenous fistula failure. The goal of our study is to investigate the influence of different angles of side-to-end radiocephalic anastomosis on the hemodynamic parameters that promote intimal dysfunction and therefore intimal hyperplasia. METHODS: Realistic three-dimensional meshes were reconstructed using ultrasound measurements from distal side-to-end radiocephalic fistulas. The velocity at the proximal and distal radial inflows and at specific locations along the anastomosis and cephalic vein was measured through duplex ultrasound performed by a single examiner. A computational parametric study, virtually changing the inner angle of anastomosis, was performed. For this purpose, we used advanced computational models that include suitable tools to capture the pulsatile and turbulent nature of the blood flow found in arteriovenous fistulas. The results were analyzed in terms of velocity fields, wall shear stress distribution, and oscillatory shear index. RESULTS: Results show that the regions with high oscillatory shear index, which are more prone to the development of hyperplasia, are greater and progressively shift toward the anastomosis area and the proximal vein segment with the decrease of the inner angle of anastomosis. These results are specific to distal radiocephalic fistulas because they are subject to proximal and distal radial inflow. CONCLUSIONS: The results of this study show that inner anastomosis angles approaching 60-70° seem to yield the best hemodynamic conditions for maturation and long-term patency of distal radiocephalic fistulas. Inner angles greater than 90°, representing the smooth loop technique, did not show a clear hemodynamic advantage.

Biopreservation as a potential hurdle for Bacillus cereus growth in fresh cheese.

This study aimed to evaluate the possible inhibitory effect of natural lactic acid bacteria on the growth of 2 Bacillus cereus strains. First, we evaluated the behavior of spores of B. cereus GPe2 and D43 when inoculated before cheesemaking using pasteurized or raw milk; no statistical differences were observed between cheese produced with the 2 types of milk. Then, lactic acid bacteria (LAB) were isolated from cheese at the last sampling time, identified, and tested in vitro for their antagonistic activity and organic acid production by using an HPLC method, showing antimicrobial potential. The LAB that produced larger inhibition halos (>9 mm) against B. cereus strains (LAB 3, 6, 9, 10: Lactococcus lactis ssp. lactis; LAB 7: Lactococcus lactis ssp. cremoris) were selected to produce a LAB mixture for subsequent tests. Spores of B. cereus GPe2 and D43 were inoculated in pasteurized milk before cheesemaking with or without addition of the LAB mixture at a high dosage. Bacillus cereus grew more slowly when LAB were added to the dairy matrix (with differences from 2.36 to 2.66 log cfu/g in B. cereus GPe2 and D43 growth).

Assessing the Disturbed Flow and the Transition to Turbulence in the Arteriovenous Fistula.

The arteriovenous fistula (AVF) is the main form of vascular access for hemodialysis patients, but its maintenance is very challenging. Its failure is mainly related to intimal hyperplasia (IH), leading to stenosis. The aim of this work was twofold: (i) to perform a computational study for the comparison of the disturbed blood dynamics in different configurations of AVF and (ii) to assess the amount of transition to turbulence developed by the specific geometric configuration of AVF. For this aim, we reconstructed realistic three-dimensional (3D) geometries of two patients with a side-to-end AVF, performing a parametric study by changing the angle of incidence at the anastomosis. We solved the incompressible Navier-Stokes equations modeling the blood as an incompressible and Newtonian fluid. Large eddy simulations (LES) were considered to capture the transition to turbulence developed at the anastomosis. The values of prescribed boundary conditions are obtained from clinical echo-color Doppler (ECD) measurements. To assess the disturbed flow, we considered hemodynamic quantities such as the velocity field, the pressure distribution, and wall shear stresses (WSS) derived quantities, whereas to quantify the transition to turbulence, we computed the standard deviation of the velocity field among different heartbeats and the turbulent kinetic energy.

Predicting growth of Listeria monocytogenes in fresh ricotta.

Challenge tests with eight brands of fresh ricotta showed rapid growth of Listeria monocytogenes and significant variability in physical-chemical characteristics. Thus, two cardinal parameters models were developed for the growth of L. monocytogenes in ricotta including, respectively, terms for temperature (Model 1) and temperature and pH (Model 2). Also an extensive, existing growth model including the effect of organic acids (Model 3) was product recalibrated to predict growth of L. monocytogenes in ricotta. Interestingly, a lack of anti-listerial effect of organic acids in ricotta was observed in this study. The range of applicability of Models 1 and 2 in ricotta (characterized by absence of competitive microbiota) included storage at temperatures from 4.1 to 20.6 °C, pH from 5.5 to 6.6, moisture contents from 72% to 82%, NaCl from 0.38% to 0.60%, concentrations of acetic acid from 579 to 1559 ppm in the water phase, citric acid from 14,774 to 46,116 ppm in the water phase, and lactic acid from 0 to 4146 ppm in the water phase. Comparing observed and predicted maximum specific growth rates of L. monocytogenes in ricotta showed a bias-factor significantly above 1, for existing models developed for broth and these models were thus fail-safe with predicted growth being faster than observed, while typically below 1 for models developed for other food types. The models developed in the present study showed bias-factors of 1.10, 1.06 and 0.78, respectively, for Model 1, 2, and 3. In particular, Model 1 and 2 developed and successfully validated could allow an easy determination of safe shelf-life of ricotta and facilitated the reformulation the product with the aim to increase shelf-life or safety.

Wet bone-in ageing and effect on beef quality technological parameters.

This study aimed to evaluate the effect of wet ageing on chemical-physical characteristics of sirloin and bone-in rib eye steaks obtained from 18 months old young bulls and 27 months old heifers. A positive beef tenderization effect of wet ageing was observed with lower values constantly detected in the rib-eye samples and significantly lower shear force values showed in heifer steaks if compared to bulls. The pH values detected both in pre- and post-ageing samples fell in the normal range for meats. The water holding capacity of sirloin samples increased with ageing while in rib eye steaks, no significant differences were detected among samples taken before and after ageing. Meat ageing resulted in a decrease of cooking loss, while meat colour parameters were not influenced by ageing and fell within the normal range. An evident improvement of the sensorial quality of bone-in wet aged beef, especially in tenderness, was achieved.

Fate of the microbial population and the physico-chemical parameters of "Sanganel" a typical blood sausages of the Friuli, a north-east region of Italy.

In Friuli, a Northeastern region of Italy, a blood sausage called Sanganel is produced by farmers, butchers, shops, and factories. This sausage is made with pork meat, boiled blood, lard, spices, and salt. It is stored at 4 ± 2 °C and usually eaten fresh or boiled within 14 days of its manufacture. Little is known about its microbial populations and safety for consumption. The aim of this study is to characterise the microbial populations and the physico-chemical parameters of Sanganel to establish its quality and the safety of consuming it. The microbial population of Sanganel is typical of meat products, and psychrotrophic enterobacteria and lactic acid bacteria (LAB) grow while it is stored. Enterobacteria produce total basic volatile nitrogen (TVB-N) and biogenic amines that, despite the presence of LAB, increase the pH of the sausage to approximately 6.9. Considering the concentrations of Enterobacteriaceae and TVB-N in the sausage, a shelf-life of 14 days is suggested. However, at 30 days the sausage is safe to eat and presents normal odours and flavours. In addition, boiling the sausage for 30 min before consumption eliminates the asporogenous microbial population.

Prevalence and quantification of thermophilic Campylobacter spp. in Italian retail poultry meat: Analysis of influencing factors.

Retail poultry meat is a crucial vehicle for consumers' exposure to Campylobacters, but no official controls are currently applied in Italy. The aim of this study was the evaluation of Campylobacter contamination of a wide range of poultry meats marketed in Italy. N. 472 chicken and turkey meat samples (sectioned meats, offal, meat preparations and products) were taken from slaughterhouses, deboning plants and different retailers and submitted to detection/enumeration of Campylobacter spp. The isolates were identified by phenotypic and biomolecular techniques. Campylobacter spp. was detected in 34.1% of the samples, with general low counts. Higher values were observed in offal (especially liver) and sectioned meats, with significantly higher rates in skin-on samples (86.8% vs 32.7%). Minced meat preparations showed lower prevalence (22.4% vs 58.3%) and counts than whole pieces. Decreasing rates were observed among slaughterhouses (80%), deboning plants (49%), butcher's shops (37%) and large scale retailers (25%). Sectioned chicken meats were significantly more contaminated than turkey meats. Almost all the isolates were identified as C. jejuni or C. coli, with similar prevalences (18.4% and 20.5%, respectively); C. jejuni was predominant only in samples from slaughterhouses/deboning plants. For setting future control programs, meat typology should be considered the main critical factor.

Microbiological and physicochemical profile of Italian steak tartare and predicting growth potential of Listeria monocytogenes.

In the present study, growth potential of Listeria monocytogenes in steak tartare samples taken at retail and belonging to 13 brands marketed in Northern Italy was investigated. The samples were submitted to microbiological and chemical-physical characterization. The data obtained were used as inputs for the application of the predictive microbiology software FSSP that allows the estimation of the growth of L. monocytogenes during the shelf-life. Lactic acid bacteria, the main component of the microflora, gave variable counts among the brands (from 3.38 to 6.24 log CFU/g). pH and aw values were always higher than 5.3 and 0.96, respectively, thus they could not be considered as single efficient hurdles to prevent the growth of L. monocytogenes according to the EC Reg. 2073/2005; the same was observed for salt content (constantly <2 %) and nitrites (not quantifiable in all the samples, even if declared in some labels). Nevertheless, the combination of all the hurdles, evaluated by predictive microbiology using critical development factors, resulted in an estimated growth <0.5 log CFU/g throughout the shelf life; this output allowed us to consider all the steak tartare analysed as unfavourable substrate for L. monocytogenes growth. The information obtained could be useful for tartare producers as well as for competent authority to evaluate the effective risk concerning these typology of products.

Leuconostoc gelidum Is the Major Species Responsible for the Spoilage of Cooked Sausage Packaged in a Modified Atmosphere, and Hop Extract Is the Best Inhibitor Tested.

Cooked sausages packaged in a modified atmosphere (MAP: 20% CO2, 70% N2, <0.2% O2) with evident yellow stains were analyzed. The aims of this work were to study the microbial cause of the spoilage and to evaluate different antimicrobial compounds to prevent it. Leuconostoc gelidum was identified as the primary cause of the yellow coating in spoiled cooked sausage, as confirmed by its intentional inoculation on slices of unspoiled sausage. Leuconostoc gelidum was the main bacteria responsible for the yellow coating in spoiled cooked sausage, as confirmed by its intentional inoculation on slices of unspoiled sausage. The yellow color was also evident during growth in the model system containing cooked sausage extract, but the colonies on MRS agar appeared white, demonstrating that the food substrate stimulated the production of the yellow pigment. The spoilage was also characterized by different volatile compounds, including ketones, ethanol, acetic acid, and ethyl acetate, found in the spoiled cooked sausage packages. These compounds explained the activity of Leuc. gelidum because they are typical of heterofermentative LAB, cultivated either on food substrates or in artificial broths. Leuc. gelidum also produced slight swelling in the spoiled packages. The efficacy of different antimicrobials was assessed in model systems composed of cooked sausage extract with the antimicrobials added at food product concentrations. The data showed that sodium lactate, sodium acetate, and a combination of sodium lactate and sodium diacetate could only slow the growth of the spoiler-they could not stop it from occurring. Conversely, hop extract inhibited Leuc. gelidum, showing a minimal inhibitory concentration (MIC) of approximately 0.008 mg CAE/mL in synthetic broth and 4 mg CAE/kg in cooked sausage slices. Adding hop extract at the MIC did not allow Leuc. gelidum growth and did not change the sensorial characteristics of the cooked sausages. To our knowledge, this is the first report of the antimicrobial activities of hop extracts against Leuc. gelidum either in vitro or in vivo.

Personalized computational electro-mechanics simulations to optimize cardiac resynchronization therapy.

In this study, we present a computational framework designed to evaluate virtual scenarios of cardiac resynchronization therapy (CRT) and compare their effectiveness based on relevant clinical biomarkers. Our approach involves electro-mechanical numerical simulations personalized, for patients with left bundle branch block, by means of a calibration obtained using data from Electro-Anatomical Mapping System (EAMS) measures acquired by cardiologists during the CRT procedure, as well as ventricular pressures and volumes, both obtained pre-implantation. We validate the calibration by using EAMS data coming from right pacing conditions. Three patients with fibrosis and three without are considered to explore various conditions. Our virtual scenarios consist of personalized numerical experiments, incorporating different positions of the left electrode along reconstructed epicardial veins; different locations of the right electrode; different ventriculo-ventricular delays. The aim is to offer a comprehensive tool capable of optimizing CRT efficiency for individual patients. We provide preliminary answers on optimal electrode placement and delay, by computing some relevant biomarkers such as d P / d t max , ejection fraction, stroke work. From our numerical experiments, we found that the latest activated segment during sinus rhythm is an effective choice for the non-fibrotic cases for the location of the left electrode. Also, our results showed that the activation of the right electrode before the left one seems to improve the CRT performance for the non-fibrotic cases. Last, we found that the CRT performance seems to improve by positioning the right electrode halfway between the base and the apex. This work is on the line of computational works for the study of CRT and introduces new features in the field, such as the presence of the epicardial veins and the movement of the right electrode. All these studies from the different research groups can in future synergistically flow together in the development of a tool which clinicians could use during the procedure to have quantitative information about the patient's propagation in different scenarios.

Shelf-life of sheep arrosticini packaged in protective atmosphere.

The aim of this study was to evaluate the shelf-life of pre-packaged sheep's arrosticini produced in a factory located in northern Italy. Samples were divided into two series and packaged in modified atmosphere with specific gas mixtures: conventional (C: 35% O2/15% CO2/50% N2) and experimental (E: 30% CO2/70% N2). All the samples were stored at 4°C for 10 days and subjected, after 5, 8 and 10 days (t5, t8, t10), to triplicate microbiological and chemical- physical (pH, total volatile basic nitrogen, thiobarbituric acid reactive substances) analyses. At the same time, the colorimetric analysis and a sensorial evaluation were carried out (pack tightness, presence of color or odor alterations), assigning a discrete score from 0 to 5. The samples showed initial total bacterial counts close to 5 Log CFU/g: these values gradually increased during storage, exceeding the threshold of 8 Log CFU/g in the C series; lower values were detected in the E series. A similar trend was highlighted for Enterobacteriaceae, with initial loads of about 3 Log CFU/g and an increase until t10, reaching values higher than 6 Log CFU/g in the C series and close to 5 Log CFU/g in the E series (P=0.002). E. coli also showed a similar trend, although with values approximately 1 Log lower than Enterobacteriaceae. Pseudomonas spp. showed initial counts close to 4.5 Log CFU/g, with a different increase in the C series (6.5 Log CFU/g at t10) and the E series (4.95 Log CFU/g) (P= 0.006). The higher growth in the C series was also observed for lactic acid bacteria, with an increase from 3 to 5 Log CFU/g (3.8 Log CFU/g in the E series P=0.016). The other microbiological parameters showed very low counts and, in most cases undetectable counts (<2 Log CFU/g) for the entire period considered. The initial values obtained from the measurement of the colorimetric indices were within the norm for this type of product: however, starting from t5, lower values of red index and lightness in the E series were measured, with an evident greying of meat surface. The results of the sensory evaluation indicated that the product showed optimal sensorial characteristics up to 8 days of shelf-life in the C series, while the use of an oxygenfree atmosphere, despite having a moderate inhibiting effect on the microbial populations, has led to an early modification of the product (5 days of storage), due to the presence of superficial greyish areas. The microbiological characteristics of arrosticini strictly depend on the hygienic conditions of slaughtering and production; even in optimal situations, the product is particularly perishable, and requires careful management of storage temperatures and times, to maintain its quality characteristics.

Prevalence, Virulence Potential, and Growth in Cheese of Bacillus cereus Strains Isolated from Fresh and Short-Ripened Cheeses Sold on the Italian Market.

This study investigated B. cereus presence in 122 samples belonging to 34 typologies of fresh or short-ripened cheeses made from cow, sheep, goat, or buffalo pasteurized milk, and sold on the Italian market. B. cereus was isolated at a prevalence of 9.8%, with a marked variability among cheese categories, and at low counts (always below 2.26 Log CFU/g). Twelve isolates were identified by MALDI-TOF analysis and typified by RAPD PCR as belonging to different B. cereus strains. All the strains were tested for the production of hemolysin BL, phosphatidylcholine-specific phospholipase C, proteases, and biofilm formation, and for the presence of chromosomal toxin-encoding genes (sph, plcA, cytK, entFM, bcet, nheA, nheB, nheC). Overall, 92% of strains harbored bcet, 75% the three genes nheA, nheB, and nheC, as well as plcA and sph, 67% entFM, and 33% cytK. All strains showed biofilm-forming ability. A chemical-physical characterization of the cheeses was also performed to show their suitability as substrates for B. cereus growth, showing high heterogeneity in terms of pH, aw, salt content, and concentration of organic acids. Finally, the ability to support spore germination and vegetative cell growth of a selected cheese was investigated in spores-inoculated samples maintained at 10 °C and 15 °C, showing the inhibitory effect of low storage temperatures.

Compliance between Food and Feed Safety: Eight-Year Survey (2013-2021) of Aflatoxin M1 in Raw Milk and Aflatoxin B1 in Feed in Northern Italy.

Aflatoxins (AFs) are fungal metabolites that are found in feed and food. When ruminants eat feed contaminated with aflatoxin B1 (AFB1), it is metabolised and aflatoxin M1 (AFM1) is excreted in the milk. Aflatoxins can result in hepatotoxic, carcinogenic, and immunosuppressive effects. The European Union thus set a low threshold limit (50 ng/L) for presence of AFM1 in milk. This was in view of its possible presence also in dairy products and that quantification of these toxins is mandatory for milk suppliers. In the present study, a total of 95,882 samples of whole raw milk, collected in northern Italy between 2013 and 2021, were evaluated for presence of AFM1 using an ELISA (enzyme-linked immunosorbent assay) method. The study also evaluated the relationship between feed materials collected from the same farms in the same area during the same period (2013-2021) and milk contamination. Only 667 milk samples out of 95,882 samples analysed (0.7%) showed AFM1 values higher than the EU threshold limit of 50 ng/L. A total of 390 samples (0.4%) showed values between 40 and 50 ng/L, thus requiring corrective action despite not surpassing the regulatory threshold. Combining feed contamination and milk contamination data, some feedingstuffs seem to be more effective in defying potential carryover of AFs from feed to milk. Combining the results, it can be concluded that a robust monitoring system that covers both feed, with a special focus on high risk/sentinel matrices, and milk is essential to guarantee high quality and safety standards of dairy products.

Evaluation of beef in purified sea water: microbiological and chemical-physical aspects.

In this study the influence of the maintenance of beef (Bavarian heifer meat submitted to dry aging for 1 month) in purified sea water was evaluated considering microbiological and physical-chemical parameters. The treated samples were perforated on their upper and lower surfaces, subsequently placed in sterile plastic containers, and covered with purified sea water while the control samples were wrapped in a plastic film; all the samples were stored at 2°C and sample analyses were performed on the day of receipt (t0) and after five (t5), seven (t7) and ten (t10) days of storage. The initial bacterial population was close to 6 log CFU/g, as expected in meat subjected to a long ageing. After 5 and 10 days of storage the treated samples showed significantly lower counts for most parameters (Total Bacterial Count, Enterobacteriaceae, Lactic Acid Bacteria and Pseudomonas spp.) if compared to the control samples. Halotolerant bacteria, yeasts and moulds showed no differences between the two series. During the test, the exposed surface of treated meat samples underwent a partial protein denaturation. Analyses show a moderate acidification of the meat during the first part of storage (from t0 to t5), followed by a substantial stabilization in the second part of the trial. The meat maintained in purified sea water acquired a significantly and constantly higher water retention due to the absorption of salt by the meat. No significant difference was detected in terms of tenderness between the two series.

Collection and analysis of post mortem inspection outcomes (liver lesions) from different cattle slaughtering plants located in Northern and Southern Italy.

The study evaluated the outcomes of cattle liver postmortem examination (64,766 animals) obtained from three slaughterhouses (two located in Lombardy, Northern Italy, and one in Puglia, Southern Italy) through the period 2016-2020. The frequency of specific lesions determining liver condemnation was calculated and the influence of several factors (animal age/category, geographical area, season, plant) was considered. A mean prevalence of 8% was observed, with a significant difference among the plants (range 6.4-12.8%). A significant difference was observed among the animal categories and age classes, with higher condemnation rates in animals aged more than 30 months (mainly cows). Steatosis was the most frequent lesion observed in cows (about half of the total), whereas liver abscesses were the most diffused in younger animals (half of the total in young bulls). Other frequently observed lesions were distomatosis and perihepatitis. A different pattern was observed between the two geographical areas, with a higher prevalence of steatosis in Lombardy (mainly intensive dairy farming), and of distomatosis in Puglia (animals mainly grazed on pasture). The season influenced the prevalence of lesions, and especially of steatosis (higher in summer). A different lesion prevalence was also observed between the two plants on similar animal populations, suggesting a difference in the notification and classification procedure among the plants. This study highlights the importance of a proper sharing procedure of the information obtained from the post mortem inspection in order to facilitate an optimal use of Food Chain Information and a useful feedback for farmers.

Bacillus cereus in Dairy Products and Production Plants.

Spore-forming Bacillus cereus is a common contaminant of dairy products. As the microorganism is widespread in the environment, it can contaminate milk at the time of milking, but it can also reach the dairy products in each phase of production, storage and ripening. Milk pasteurization treatment is not effective in reducing contamination and can instead act as an activator of spore germination, and a potential associated risk still exists with the consumption of some processed foods. Prevalences and concentrations of B. cereus in milk and dairy products are extremely variable worldwide: in pasteurized milk, prevalences from 2% to 65.3% were reported, with concentrations of up to 3 × 105 cfu/g, whereas prevalences in cheeses ranged from 0 to 95%, with concentrations of up to 4.2 × 106 cfu/g. Bacillus cereus is also well known to produce biofilms, a serious concern for the dairy industry, with up to 90% of spores that are resistant to cleaning and are easily transferred. As the contamination of raw materials is not completely avoidable, and the application of decontamination treatments is only possible for some ingredients and is limited by both commercial and regulatory reasons, it is clear that the correct application of hygienic procedures is extremely important in order to avoid and manage the circulation of B. cereus along the dairy supply chain. Future developments in interventions must consider the synergic application of different mild technologies to prevent biofilm formation and to remove or inactivate the microorganism on the equipment.

Computational electrophysiology of the coronary sinus branches based on electro-anatomical mapping for the prediction of the latest activated region.

This work dealt with the assessment of a computational tool to estimate the electrical activation in the left ventricle focusing on the latest electrically activated segment (LEAS) in patients with left bundle branch block and possible myocardial fibrosis. We considered the Eikonal-diffusion equation and to recover the electrical activation maps in the myocardium. The model was calibrated by using activation times acquired in the coronary sinus (CS) branches or in the CS solely with an electroanatomic mapping system (EAMS) during cardiac resynchronization therapy (CRT). We applied our computational tool to ten patients founding an excellent accordance with EAMS measures; in particular, the error for LEAS location was less than 4 mm. We also calibrated our model using only information in the CS, still obtaining an excellent agreement with the measured LEAS. The proposed tool was able to accurately reproduce the electrical activation maps and in particular LEAS location in the CS branches, with an almost real-time computational effort, regardless of the presence of myocardial fibrosis, even when information only at CS was used to calibrate the model. This could be useful in the clinical practice since LEAS is often used as a target site for the left lead placement during CRT. Overall picture of the computational pipeline for the estimation of LEAS.

Screening of species-specific lactic acid bacteria for veal calves multi-strain probiotic adjuncts.

The selection of promising specific species of lactic acid bacteria with potential probiotic characteristics is of particular interest in producing multi species-specific probiotic adjuncts in veal calves rearing. The aim of the present work was to select and evaluate in vitro the functional activity of lactic acid bacteria, Bifidobacterium longum and Bacillus coagulans strains isolated from veal calves in order to assess their potential use as multi species-specific probiotics for veal calves. For this purpose, bacterial strains isolated from faeces collected from 40 healthy 50-day-calves, were identified by RiboPrinter and 16s rRNA gene sequence. The most frequent strains belonged to the species B. longum, Streptococcus bovis, Lactobacillus animalis and Streptococcus macedonicus. Among these, 7 strains were chosen for testing their probiotic characteristics in vitro. Three strains, namely L. animalis SB310, Lactobacillus paracasei subsp. paracasei SB137 and B. coagulans SB117 showed varying individual but promising capabilities to survive in the gastrointestinal tract, to adhere, to produce antimicrobial compounds. These three selected species-specific bacteria demonstrated in vitro, both singularly and mixed, the functional properties needed for their use as potential probiotics in veal calves.

Growth potential of Listeria monocytogenes in veal tartare.

In the present study the growth potential of Listeria monocytogenes in veal tartare was evaluated. A challenge test was performed on three tartare batches at 8°C, aiming to evaluate the growth potential of the pathogen. The data indicated the absence of a significant growth (δ<0.5 log cfu/g) during the entire period. When considering intermediate sampling times, an increase of 0.56 log cfu/g was detected after five days of storage in one of the batches. Microflora of veal tartare was dominated by lactic acid bacteria, that increased gradually during the trial, reaching counts up to 7 Log CFU/g in two of the three batches considered. Spoilage bacteria were present (especially Pseudomonas spp., yeasts and Enterobacteriaceae) but in very low counts and with a limited increase during the period considered. Finally, daily maximum tolerable L. monocytogenes counts were calculated to highlight the maximum acceptable load to avoid the overcoming of the legal limit of 100 CFU/g: a total increase of 0.95 log cfu/g in 12 days of shelf-life was estimated, obtaining a "safety initial concentration" at t0 of 10 CFU/g of the pathogen.