Enhancing the quality and safety of Nocellara del Belice green table olives produced using the Castelvetrano method.

The Castelvetrano method is the most widely used among the various table olive processing styles in Sicily. After debittering, the product is stored at low temperatures to prevent the growth of undesirable microorganisms. In an effort to enhance the production process, yeast isolates underwent genotypic characterization and technological screening. The screening process identified two yeast strains Candida norvegica OC10 and Candida boidinii LC1, which can grow at low temperatures and tolerate high pH values (up to 10) and salinity [10% (w/v)]. During the monitoring period, the inoculated trials showed limited presence of spoilage/pathogenic microorganisms. Additionally, the yeasts limited oxidative phenomena and softening of the drupes. The organic compounds detected were higher in the inoculated trials than in the control, and cold storage induced aromatic decay, which was less pronounced in the trial inoculated with C. norvegica. Sensory analysis revealed that the inoculated trials scored higher in sweetness, hardness and crispness.

Evaluation of different conditions to enhance the performances of Lactobacillus pentosus OM13 during industrial production of Spanish-style table olives.

The main objective was to set up a methodology to improve the high volume production of green table olives, cv. Nocellara del Belice. Lactobacillus pentosus OM13 was applied during three different industrial processes of table olives as follows: trial one (IOP1) was subjected to an addition of lactic acid until a brine level of pH 7.0 was reached; trial two (IOP2) subjected to same addition of lactic acid as in trial one plus nutrient adjuvant and trial three (IOP3) subjected to same addition as trial two, but with the strain L. pentosus OM13 acclimatized in brine for 12 h before inoculation. These trials were compared against two untreated controls (spontaneously fermented and addition of L. pentosus OM13 only). Within the third day of fermentation, the pH of the brines decreased significantly, reaching pH 4.85 for trial three, pH 5.15 for trial two, and pH 5.92 for trial one. The pH of both controls decreased more slowly, and had values below pH 5.0 only after the fifteenth day of fermentation (control one) and the sixty-fifth day of fermentation (control two). Trial three reached the highest lactic acid bacteria (LAB) concentration on the third day of fermentation. After six days of fermentation, all trials showed similar values of LAB counts that were significantly higher compared to control number one. The result from genotypic identification showed that L. pentosus OM13 was the most frequently isolated in the inoculated trials. Lactobacillus plantarum, Lactobacillus coryniformis and Pediococcus pentosaceous were also detected at very low concentrations. Homoguaiacol, 2-butanol, 4-ethylphenol, phenylethyl alcohol and 4-ethylphenol were the volatile organic compounds detected at the highest levels in all experimental trials. Trial three showed a higher concentration of squalene that was not detected in other trials. The highest sensory scores of green olive aroma and overall satisfaction were found for all experimental olives, especially for those of trial one and trial two, that differed significantly from the untreated controls. This study provides evidence that the addition of lactic acid, nutrient adjuvants and, most importantly, the acclimatization of LAB cells significantly shortens the acidification process of olive brine, and improves safety and sensory quality. Shorter acidification processes result in a more rapid transformation of table olives, with reduced commodity loss and lower costs of production compared to conventional manufacturing protocols.

Pneumococcal colonization in older adults.

BACKGROUND: Little is known about pneumococcal carrier states in older adults. The main aim of this study was to evaluate pneumococcal colonization patterns among older adults in two centres in Milan, Italy, before the widespread use of the 13-valent pneumococcal vaccine (PCV13) in this age group, to investigate demographic and clinical features that are associated with pneumococcal colonization and to estimate the potential coverage offered by PCV13. RESULTS: Among 417 adults ≥65 years old (171, 41.1 %, ≥75 years), 41 (9.8 %) were pneumococcal carriers. Univariate and multivariate analyses revealed that pneumococcal colonization was significantly less common among individuals with underlying co-morbidities than among those without (odds ratio [OR] 0.453, 95 % confidence interval [CI] 0.235-0.875, p = 0.018; adjusted OR 0.503, 95 % CI 0.255-0.992, p = 0.047). Moreover, among these patients, those with cardiac disease had a significantly lower risk of colonization (OR 0.308, 95 % CI 0.119-0.795, p = 0.015; adjusted OR 0.341, 95 % CI 0.13-0.894, p = 0.029). Only one vaccinated subject who received 23-valent polysaccharide pneumococcal vaccine (PPV23) was colonized. Twenty-five (89.3 %) of the subjects who were <75 years old and 9 (75.0 %) of those who were ≥75 years old were colonized by at least one of the serotypes that is included in PCV13, with serotype 19 F being the most common. Respiratory allergies as well as overall co-morbidities were more common in subjects who were positive for only non-PCV13 serotypes compared with negative subjects and those who were carriers of only PCV13 serotypes. CONCLUSIONS: Although this study included a relatively small number of subjects and has been performed in a limited geographic setting, results showed that pneumococcal colonization in older people is common, and the monitoring of carriers can offer useful information about the circulation of this pathogen among older people and the potential protective effect of pneumococcal vaccines. Because the colonization in most cases involves the strains that are included in PCV13, this vaccine could be useful in the prevention of pneumococcal infections in the overall population of older people. In subjects with respiratory allergies and in those with co-morbidities, the addition of the PPV23 to PCV13 should be recommended. Due to the low vaccination coverage, urgent educational programmes are required to inform older adults and their medical doctors of the risks of pneumococcal infection and the efficacy and safety of the available pneumococcal vaccines.

Effect of the mechanical harvest of drupes on the quality characteristics of green fermented table olives.

BACKGROUND: Because of damage caused by mechanical harvesting, the drupes for table olive production are traditionally hand harvested. Until now, no data have been available on the microbiological and chemical features of mechanically harvested drupes during fermentation. RESULTS: Drupes mechanically harvested and inoculated with Lactobacillus pentosus OM13 were characterized by the lowest concentrations of potential spoilage microorganisms. On the other hand, drupes mechanically harvested and subjected to spontaneous fermentation showed the highest concentration of Enterobacteriaceae and pseudomonads during transformation. The lowest decrease of pH (4.20) was registered for the trials inoculated with the starter culture. Differences in terms of volatile organic compounds were estimated among trials. Multivariate analysis showed that the olives processed from the drupes mechanically harvested and inoculated with starter were closely related to control production (drupes manually harvested) in terms of microbiological and pH values. Sensory analysis evidenced negative evaluations only for the uninoculated trials. CONCLUSION: Drupes mechanically harvested and subjected to a driven fermentation with Lactobacillus pentosus OM13 determined the production of table olives with appreciable organoleptic features. Thus mechanical harvesting performed using a trunk shaker equipped with an inverse umbrella and the addition of starter lactic acid bacteria represents a valuable alternative to manual harvesting for table olive production at the industrial level.

An innovative method to produce green table olives based on "pied de cuve" technology.

The technology of "pied de cuve" (PdC) is applied in food process only to produce wines with an enriched community of pro-technological yeasts. PdC promotes the growth of the desirable microbial strains in a small volume of grape must acting as a starter inoculums for higher volumes. The aim of the present work was to investigate the use of partially fermented brines, a technology known as PdC, developed with lactic acid bacteria (LAB) on the microbiological, chemical and sensory characteristics of green fermented table olives during two consecutive campaigns. The experimental plan included two trials based on different PdCs: trial A, PdC obtained with Lactobacillus pentosus OM13; trial B, PdC obtained through a spontaneous fermentation. Two control additional trials without PdC were included for comparison: trial C, spontaneous fermentation; trial D, direct inoculation of L. pentosus OM13. The use of PdCs favoured the rapid increase of LAB concentrations in both trials A and B. These trials showed levels of LAB higher than trial C and almost superimposable to that of trial D. Trial B was characterized by a certain diversity of L. pentosus strains and some of them dominated the manufacturing process. These results indicated PdC as a valuable method to favour the growth of autochthonous L. pentosus strains. Hierarchical cluster analysis (HCA) and principal component analysis (PCA) visibly discriminated olive processes fermented with the two experimental PdCs. Interestingly, on the basis of microbial and pH variables, both approaches showed that the olives produced with PdC technology are closely related to those of trial D, with the advantages of reducing the amount of starter to inoculate (trial A) and a higher LAB biodiversity (trial B). Volatile organic compound (VOC) composition and sensory analysis showed trials A and B different from the trials with no PdC added, in both years. Furthermore, the trial B showed the highest scores of green olive aroma and taste complexity. Spoilage microorganisms were estimated at very low levels in all trials. Undesired off-odours and off-flavours were not revealed at the end of the process.

Use of Different Nutrients to Improve the Fermentation Performances of Lactiplantibacillus pentosus OM13 during the Production of Sevillian Style Green Table Olives.

The aim of this study was to evaluate the fermentation performance of the commercial starter Lactiplantibacillus pentosus OM13 with four nutrients (A, B, C, and D) that differed in the following ingredients: starch, sugars, maltodextrin, inactivated yeast, inactivated yeast rich in amino acids, inactivated yeast rich in mannoproteins, and salt (NaCl). For this purpose, six different experimental productions of Nocellara del Belice table olives were carried out. During transformation, the fermentation process was monitored by measuring pH and plate counts for lactic acid bacteria (LAB), yeasts, Enterobacteriaceae, Staphylococcaceae, and Pseudodomondaceae populations. At the end of the production process, each trial was subjected to volatile organic compound analysis and sensory evaluation. The addition of the different nutrients resulted in a significant reduction in pH (around 2.5 points) after 3 days of fermentation. At the same time, a significant increase in the number of LAB populations (> 6.6 log CFU/mL) was observed for all trials. Volatile organic compound (VOC) analysis revealed the presence of 39 compounds. In this study, nutrient C was optimal for improving the fermentation activity of L. pentosus OM13. These results provide elements for the implementation of experimental protocols to reduce product losses and improve sensory characteristics.

Use of selected autochthonous lactic acid bacteria for Spanish-style table olive fermentation.

The present work presents a successful attempt to achieve an enhanced and more predictable fermentation process in Spanish-style green olive technology by selection and use of autochthonous starter cultures. During the first phase of this work, two Spanish-like fermentations of green table olives of cultivar (cv) "Nocellara del Belice", coming from irrigated and not irrigated fields, were monitored, in order to highlight the best agricultural conditions for drupe production and to isolate lactic acid bacteria strains with relevant technological properties. Among 88 identified isolates, one Lactobacillus pentosus strain showed remarkable biochemical features and high acidification rate in synthetic brine. In the second phase, the selected strain was used as starter culture in three different trials to establish the best conditions for its use. Microbial counting, as well as starter tracking by M13 RAPD-PCR, reflected the optimal adaptation of the strain to the environment. Spontaneous fermentation needed a 14-day long lag phase to reach the same population as the inoculated trials. Moreover, sensory traits of table olives obtained with adjunct culture showed better characteristics compared to those processed in the other trials, in particular concerning the presence of off-odours.

Influenza vaccination among healthcare workers in a multidisciplinary University hospital in Italy.

BACKGROUND: Annual influenza vaccination is recommended for healthcare workers (HCWs) in order to reduce the morbidity associated with influenza in healthcare settings. The aim of this study was to evaluate the current vaccination status of the HCWs in one of Italy's largest multidisciplinary University Hospitals. METHODS: Between February 1 and March 31, 2006, we carried out a cross-sectional study of influenza vaccination coverage among HCWs at the University Hospital Fondazione IRCCS "Ospedale Maggiore Policlinico, Mangiagalli e Regina Elena", Milan, Italy. After receiving a brief description of the aim of the study, 2,143 (95%: 1,064 physicians; 855 nurses; 224 paramedics) of 2,240 HCWs self-completed an anonymous questionnaire. RESULTS: Influenza vaccination coverage was very low in all specialties, varying from 17.6% in the Emergency Department to 24.3% in the Surgery Department, and knowledge of influenza epidemiology and prevention was poor. The factors positively associated with being vaccinated were an age of >or= 45 years, considering influenza a potentially severe disease, and being aware of the high-risk categories for which influenza vaccination is strongly recommended; those that negatively associated with being vaccinated were being female, working in the Medicine Department, and being a nurse or paramedic. CONCLUSION: Despite strong recommendations, influenza vaccination coverage seemed to be very low among HCWs of all specialties, with differences between areas and types of employment. Specific continuous educational and vaccination programs for different targets should be urgently organized to reduce morbidity and mortality in high-risk patients, contain nosocomial outbreaks, and ensure an appropriate socioeconomic impact.

Disseminated Penicillium marneffei infection in an HIV-positive Italian patient and a review of cases reported outside endemic regions.

We describe a case of disseminated Penicillium marneffei in a human immunodeficiency virus (HIV)-positive Italian man who stayed for 4 years in Chiang Ray province, northern Thailand. A review of the literature shows that penicilliosis, although unusual, may represent an emerging opportunistic infection among HIV-positive people traveling to endemic areas.

Unusual codon 69 insertions: influence on human immunodeficiency virus type 1 reverse transcriptase drug susceptibility.

INTRODUCTION: Multiple amino acid changes in the reverse transcriptase (RT) enzyme of the human immunodeficiency virus 1 (HIV-1) confer simultaneous resistance to most nucleoside RT inhibitors (NRTI). It may take place through different pathways: one of these is the codon 69 insertion, which can involve several 2-amino acid patterns. MATERIALS AND METHODS: We are reporting the case of three patients treated with various antiretroviral compounds. For these subjects we have conducted both a genotypical and a phenotypical analysis in order to understand what kind of influence these insertions may have on HIV-1 RT drug susceptibility. Plasma samples from these patients have been extracted and the RT region has been amplified, cloned and sequenced; meanwhile their PBMCs have been separated, cultivated and then tested for drug susceptibility. RESULTS: Data obtained from the cloning assay showed that the patients had different mutational patterns but constant multiple resistance to NRTI. In particular, they harbored mutations related to Zidovudine (ZDV), 3TC and various NRTIs. Moreover, all three samples had a T69S substitution followed by three different dual amino acid insertions: SG, TG and VG. Several phenotypic experiments revealed that the viruses were resistant to 3TC as well as to ZDV and ABC. Different results were obtained using d4T and ddI. DISCUSSION: In our three patients, all mutation inserts impaired the use of NRTI, particularly ZDV and 3TC. Patient 001 presented a pattern that should not cause a high phenotypic resistance to 3TC per se, and so we can argue that the concomitant presence of the insertion T69S (SG) makes this isolate moderately resistant to this drug. We observed a similar phenomenon in subject 003. d4T was less involved in the resistance generation caused by the RT insertion (in one out of three cases). Moreover, we identified a new 2aa insertion (TG) that has, to the best of our knowledge, never been reported before. A careful survey of novel RT genotypic insertion is thus warranted.

Paraurethral leiomyoma.

Paraurethral leiomyoma is a rare, benign, hormone-dependent neoplasm of mesenchymal origin affecting women. The clinical evidence varies, but it tends to be asymptomatic or associated with the sensation of a foreign body; urinary symptoms are rarely described. The distinction among urethral, paraurethral, and anterior vaginal wall leiomyoma can be very difficult owing to their anatomic proximity. Excision of the mass is the recommended treatment, and the diagnosis is confirmed by the pathologic finding to rule out the presence of a sarcoma. A case of paraurethral leiomyoma associated with dysuria, dyspareunia, and obstructive voiding symptoms is reported.

Risk factors for HIV/AIDS in a low HIV prevalence site of sub-Saharan Africa.

We conducted a hospital-based survey on prevalence and risk factors of HIV-1/2 and other viral infections in Zanzibar archipelago. Blood samples, socio-demographic and behavioural data were collected from 2697 patients. The overall HIV prevalence was 2.9%. About 1.4%, 2.1%, 4.2% of antenatal clinic (ANC) attendees and 2.1%, 3.7%, 5.3% of blood donors were, respectively, HIV-Abs-, HTLV-Abs- and HBs-Ag-positive; 5.5% of blood donors were HCV-affected. Co-infections were rare. Exactly 3.4% of the children aged 6-10 years were HIV-positive. People aged 26-35 years [adjusted odds ratio (AOR) 4.4, 95% CI (confidence interval) 1.72-11.22; P = 0.002], illiterate subjects (AOR 3.6, 95% CI 1.65-7.98; P = 0.001) mobile workers (AOR 7.0, 95% CI 1.41-34.62; P = 0.02) and previously operated patients (AOR 1.9, 95% CI 1.02-3.66; P = 0.04) were at higher risk for HIV/AIDS. Any of the examined factors were associated with hepatitis B virus, hepatitis C virus and human T lymphotropic virus type 1/2 transmission. HIV/AIDS prevention strategies must primarily be addressed to traditional high-risk groups and secondarily to unsafe health care procedures in relatively preserved sub-Saharan areas.

Analysis of protease inhibitor combinations in vitro: activity of lopinavir, amprenavir and tipranavir against HIV type 1 wild-type and drug-resistant isolates.

BACKGROUND: Despite the increasing number of antiretroviral compounds, the number of useful drug regimens is limited owing to the high frequency of cross-resistance. PATIENTS AND METHODS: We studied in vitro two-drug combinations using three protease inhibitors (PIs), tipranavir, amprenavir and lopinavir, on isolates (003 and 004) derived from patients with resistance to multiple PIs compared with the drug-susceptible isolate 14aPre in peripheral blood mononuclear cells. Drug interactions were determined by median dose-effect analysis, with the combination index calculated at several inhibitory concentrations (IC). RESULTS: In 14aPre experiments, the combination tipranavir + lopinavir demonstrated synergy at low concentrations (IC(50)), an additive effect at IC(75) and antagonism at IC(90)-IC(95); tipranavir + amprenavir were antagonistic at all concentrations except IC(95), where they were synergic; and the lopinavir + amprenavir combination was always antagonistic. In 003 and 004 infections, tipranavir + lopinavir and tipranavir + amprenavir combinations were antagonistic, and lopinavir + amprenavir were synergic, at all concentrations, with the exception of being additive at IC(95). CONCLUSIONS: Our in vitro experiments did not show any advantage in combining second generation PIs as a therapeutic strategy in naive or multi-treatment failure subjects, with the exception of tipranavir + amprenavir at IC(95) in infections by a wild-type isolate.