Effectiveness of the Thermal Treatments Used for Curd Stretching in the Inactivation of Shiga Toxin-Producing O157 and O26 Escherichia coli.

The kneading treatment of the fresh curd in hot water is a critical control point in the manufacturing of mozzarella. Factors such as the ratio between hot water and curd mass, the rheological properties, and the mixing and kneading activity affect the processing time and the internal temperature of the curd. The aim of this study was to investigate the effect of thermal treatments on the fate of Shiga toxin-producing Escherichia coli (STEC). Nine curd samples (weight 160-270 g) were artificially contaminated with O157 or O26 STEC and stretched in hot water (90-95°C) for 5-10 min. Depending on the heating process and spinning, different nonisothermal profiles were recorded. Observed reductions of O157 and O26 STEC varied between 1.01 and more than 5.38 log⁡MPN (Most Probable Number)/g at the end of the temperature treatments. Further, nonisothermal log-linear tail models were developed to compare observed reductions for O157 and O26 VTEC under variable temperature conditions. Results obtained showed that the comparison of predictions provided by the dynamic model with observations described well the linear inactivation pattern since nonsignificant differences were denoted at all profiles tested. The dynamic model developed can be useful to evaluate the effectiveness of the thermal treatments used in the manufacturing of mozzarella in the inactivation of STEC.

[Prevalence of atrial fibrillation, treatment eligibility and consumption of oral anticoagulants in Italian Local Health Authorities: impact of non-vitamin K antagonist oral anticoagulants]. / Prevalenza della fibrillazione atriale, eleggibilità al trattamento e consumo di anticoagulanti orali nelle Aziende Sanitarie italiane: impatto dei nuovi anticoagulanti.

BACKGROUND: Atrial fibrillation (AF) increases significantly the risk of comorbidities and premature death. This paper aims to estimate: 1) the current prevalence of AF in Italy as a whole and within individual local health authorities (LHA); 2) the proportion of strongly eligible patients receiving oral anticoagulants; 3) the impact of non-vitamin K antagonist oral anticoagulants (NOACs). METHODS: The prevalence of AF and the proportion of patients strongly eligible for anticoagulant treatment (defined by a CHADS2 score ≥2) were obtained by applying the results of a systematic literature search to the Italian population stratified by gender, age and LHA. The consumption of oral anticoagulants in eligible patients within each LHA was derived from Intercontinental Marketing Services data. RESULTS: Altogether, the detected prevalence of AF in Italy is 1.7%, i.e. 1 036 448 cases. Of these, 62.6%, i.e. 648 832 subjects, are estimated to have a CHADS2 ≥2 and therefore strongly eligible for anticoagulant treatment. The estimated percentage of eligible treated patients in 2015 was 43.7%, significantly greater compared to 2014 (31.3%), with local and regional variabilities. In 2015 the estimated use of NOACs for AF was 36.4% of all oral anticoagulants, with marked local and regional variabilities. NOAC prescription appears influenced by financial limitations and by factors related to the network of anticoagulation services. CONCLUSIONS: The present analysis shows a prevalence of AF in Italy of 1.7%. The overall rate of anticoagulant use is increasing, but is still insufficient, with marked regional and local variations. In 2015, approximately one third of oral anticoagulation in eligible patients was based on NOACs.

Detection of Shiga toxin (Stx)-producing Escherichia coli (STEC) in bovine dairy herds in Northern Italy.

The aim of this study was to monitor the presence of Shiga toxin (Stx)-producing Escherichia coli in dairy farms authorized to sell raw milk and other farms, located in the same area, which sell milk to industry or use it to produce Parmesan or Grana cheese. Our research was focused on the serogroups O157 and O26, which are the most common in human cases in Italy and genetic markers that characterize the strains that can cause hemorrhagic colitis and hemolytic uremic syndrome (EHEC) in humans. Overall, 255 bulk-milk and 225 milk filter samples were screened for the presence of Shiga toxin genes (stx1 and stx2), O157 and O26 serogroups by using PCR. The samples were collected in 193 bovine dairy farms located in Northern Italy, including 32 farms selling raw milk to consumers. According to the preliminary PCR screening test, 32 out of 255 (12.5%; CI95%, 8.7% to 17.3%) bulk milk samples and 68 out of 225 (30.2%; CI95%, 24.3% to 36.7%) milk filters were positive for stx genes. Of the 32 milk samples that were stx-positive, 4 (1.6%, CI95%, 0.4% to 4%) were also positive by PCR for the rfbEO157 gene and 6 (2.4%, CI95%, 0.9% to 5.1%) were positive for the wzxO26 gene. The culture detection method, which was based on the immunomagnetic separation, achieved isolation rates of E. coli serogroups O157 and O26 in 25-67% of the milk samples that tested positive by PCR for these serogroups. STEC O26 was detected in one milk filter (1.6%) from a farm that sells raw milk to consumers directly and one sample (1.4%) of bulk milk intended for pasteurization. The presence of STEC O157 was also detected in 2 milk filters (1.7%) from farms that use milk to produce Grana cheese. All the STEC stains O157 and O26 isolated carried the genes eae and espK and genes belonging to the pathogenicity island OI-122 (efa1/2, sen, pagC), which are markers suitable for screening the human virulent EHEC strains. These virulence markers were also detected in the three strains of stx-negative E. coli O157 isolated from two filters and one milk sample. These strains could be therefore EHEC strains that have lost the stx genes (EHEC-derivative strains). Concern arise for the presence of EHEC O26 and E. coli O157 isolates that are suspected to be an EHEC-derivative in the milk filters sampled in farms that are used to sell raw milk to consumers and in other dairy farms.

Thermal inactivation kinetics of Shiga toxin-producing Escherichia coli in buffalo Mozzarella curd.

The use of raw milk in the processing of buffalo Mozzarella cheese is permitted, but the heat treatment used for stretching the curd must ensure that the final product does not contain pathogens such as Shiga toxin-producing Escherichia coli (STEC) that may be present on buffalo dairy farms. This study carried out challenge tests at temperatures between 68 °C and 80 °C for 2 to 10 min to simulate curd temperatures during the stretching phase. Curd samples were inoculated with 2 STEC strains (serotypes O157 and O26), and their inactivation rates were assessed in the different challenge tests. The curd samples were digested with papain to ensure a homogeneous dispersion of bacteria. The STEC cells were counted after inoculation (range 7.1-8.7 log cfu/g) and after heat treatments using the most probable number (MPN) technique. A plot of log MPN/g versus time was created for each separate experiment. The log linear model with tail was used to provide a reasonable fit to observed data. Maximum inactivation rate (k(max), min(-1)), residual population (log MPN/g), decimal reduction time (min), and time for a 4D (4-log10) reduction (min) were estimated at each temperature tested. A 4D reduction of the O26 STEC strain was achieved when curd was heated at 68 °C for 2.6 to 6.3 min or at 80 °C for 2.1 to 2.3 min. Greater resistance was observed for the O157 strain at 68 °C because k(max) was 1.48 min(-1). The model estimates can support cheesemakers in defining appropriate process criteria needed to control possible STEC contamination in raw milk intended for the production of Mozzarella.

Intraoperative detection of lung cancer by octreotide labeled to Indium-111.

The contribution of radio surgery (RGS) using octreotide labeled with Indium-111 (In111) has been studied for surgical treatment of lung cancer. Thirteen patients were administered 111 Mbq of In111 octreotide intravenously. Scintigraphic images were preoperatively taken at 4, 24, and 48 hours after the tracer injection. Pulmonary resection and intraoperative evaluation by RGS technique were then performed to set the section limits. Histological staining of all the resected specimens and resection margins were assessed and their results were used as a confirmation of the RGS intraoperative findings. RGS is a simple method that can help the surgeon in the intraoperative assessment of bronchial, parenchymal, and parietal resection margins. Further research is needed to verify whether this method also may be useful in the intraoperative definition of the extent of mediastinal lymph node dissection.

[Preliminary experience with radioimmuno-guided surgery of primary neoplasms of the lung]. / Esperienze preliminari sulla chirurgia radioimmunoguidata delle neoplasie primitive del polmone.

BACKGROUND: Radioimmunoguided surgery (RIGS) can be a valid option in the management of lung cancer as well as neoplasms in other anatomic sites. METHODS: We evaluated the usefulness of radioimmunoguided surgery (RIGS) in the staging of primitive non small cell lung cancer. Intraoperatively, this technique can define the lymph nodes involvement and thus, the radicality of the resection. In the first stage of our study, we looked for the epitope TAG 72 in 45 patients with primary non small cell lung cancer. The epitope was found by immunochemistry in only 6 cases. The only one operable patient was injected with monoclonal antibody B 72.3, that was 125I-labelled. RESULTS: At the operation, the monoclonal antibody showed no selectivity for neoplastic cells. Neoplastic tissue and healthy tissue showed a similar detection of the monoclonal antibody both intraoperatively and at the histochemical study. Because of the problems related with this method--e.g. technical difficulties, excessive wasting of time and lack of imaging--we modified our strategy. In this second stage of our study we used fragments of murine anti-CEA monoclonal antibody F023C5. The protocol was performed in 11 patients with squamous cell lung cancer. In one patient operated on for an excavated cancer(not well-defined at the immunoscintigraphy) intraoperative detection was negative while the ex vivo counts were significant: the neoplastic tissue showed a radioactivity twice higher than healthy tissue. Furthermore, the RIGS found a small intraparenchimal lymph node that was seen neither by CT nor by immunoscintigraphy. CONCLUSIONS: Our data are still preliminary, but with improvement of the technique and the use of more specific monoclonal antibodies the RIGS could become a helpful method, able to improve the radicality of surgery for lung cancer.