Cross-contamination of mature Listeria monocytogenes biofilms from stainless steel surfaces to chicken broth before and after the application of chlorinated alkaline and enzymatic detergents.

The objectives of this study were, firstly, to compare a conventional (i.e., chlorinated alkaline) versus an alternative (chlorinated alkaline plus enzymatic) treatment effectivity for the elimination of biofilms from different L. monocytogenes strains (CECT 5672, CECT 935, S2-bac and EDG-e). Secondly, to evaluate the cross-contamination to chicken broth from non-treated and treated biofilms formed on stainless steel surfaces. Results showed that all L. monocytogenes strains were able to adhere and develop biofilms at approximately the same growth levels (≈5.82 log CFU/cm2). When non-treated biofilms were put into contact with the model food, obtained an average transference rate of potential global cross-contamination of 20.4%. Biofilms treated with the chlorinated alkaline detergent obtained transference rates similar to non-treated biofilms as a high number of residual cells (i.e., around 4 to 5 Log CFU/cm2) were present on the surface, except for EDG-e strain on which transference rate diminished to 0.45%, which was related to the protective matrix. Contrarily, the alternative treatment was shown to not produce cross-contamination to the chicken broth due to its high effectivity for biofilm control (<0.50% of transference) except for CECT 935 strain that had a different behavior. Therefore, changing to more intense cleaning treatments in the processing environments can reduce risk of cross-contamination.

Dual-species biofilms formation between dominant microbiota isolated from a meat processing industry with Listeria monocytogenes and Salmonella enterica: Unraveling their ecological interactions.

Alternatives to combat the persistence of pathogens need to consider the microbiota established on industrial surfaces as they can influence the protection or replacement (i.e. reduction/inhibition) of pathogens. The objective of the present study was to determine the ecological interactions established in dual-species biofilms between Listeria monocytogenes and Salmonella enterica as target pathogens, and isolates recovered from a meat processing facility (i.e.Pseudomonas fluorescens, Pseudomonas fragi, Bacillus safensis, Bacillus megaterium, and Candida zeylanoides). Results showed different ecological relations in biofilms depending on the species evaluated. Pseudomonas spp. did not influence the growth of either pathogen, although tested species tended to protect the pathogens in the structures generated. B. megaterium and C. zeylanoides affected the two pathogens differently, demonstrating a reduction of L. monocytogenes adhered cells within the formed biofilm. B. safensis reduced or presented non-influence on S. enterica depending on the incubation conditions. Contrarily, B. safensis was the microorganism that demonstrated the highest replacement capacity for L. monocytogenes, reducing its growth by up to 4 log CFU/cm2. The in vitro study of bispecies biofilms is important for the food industry, helping to understand how they behave and to find an effective way to eliminate them.

Robust gene signatures from microarray data using genetic algorithms enriched with biological pathway keywords.

Genetic algorithms are widely used in the estimation of expression profiles from microarrays data. However, these techniques are unable to produce stable and robust solutions suitable to use in clinical and biomedical studies. This paper presents a novel two-stage evolutionary strategy for gene feature selection combining the genetic algorithm with biological information extracted from the KEGG database. A comparative study is carried out over public data from three different types of cancer (leukemia, lung cancer and prostate cancer). Even though the analyses only use features having KEGG information, the results demonstrate that this two-stage evolutionary strategy increased the consistency, robustness and accuracy of a blind discrimination among relapsed and healthy individuals. Therefore, this approach could facilitate the definition of gene signatures for the clinical prognosis and diagnostic of cancer diseases in a near future. Additionally, it could also be used for biological knowledge discovery about the studied disease.

Removal of Listeria monocytogenes biofilms on stainless steel surfaces through conventional and alternative cleaning solutions.

Conventional treatments are not effective enough to completely remove Listeria monocytogenes biofilms from surfaces, thus implying the presence of certain persistent bacterial forms. In this study, eleven treatments (i.e. two enzymatic agents applied at two different temperatures and concentrations, two alkaline cleaners and one acid detergent) were used to remove mature L. monocytogenes S2-bac biofilms. A combined treatment was then selected for its application to four different L. monocytogenes strains (i.e. CECT 5672, CECT 935, S2-bac, EDG-e). Effectivity of the treatments was evaluated quantitatively using TEMPO and qualitatively by direct epifluorescent microscopy (DEM). Bacterial detachment obtained after the application of acid, alkaline and chlorinated alkaline treatments were 6.03, 6.24 and 4.76 Log CFU/cm2, respectively. Enzymatic treatments applied at 50 °C obtained the greatest detachment and biocidal activity. The results derived from the observation of the remaining biofilm structure by DEM proved that conventional treatments were unable to completely remove conformed structures with the potential risk this entails. Last, the application of a combined treatment using a chlorinated alkaline cleaner followed by an enzymatic treatment enhanced the dispersal of the bacterial cells from surfaces, thus consolidating this as a good option to recommend for the 5-step cleaning procedure.

Electronic health records and patient registries in medical oncology departments in Spain.

PURPOSE: We aimed to evaluate the current situation of electronic health records (EHRs) and patient registries in the oncology departments of hospitals in Spain. METHODS: This was a cross-sectional study conducted from December 2018 to September 2019. The survey was designed ad hoc by the Outcomes Evaluation and Clinical Practice Section of the Spanish Society of Medical Oncology (SEOM) and was distributed to all head of medical oncology department members of SEOM. RESULTS: We invited 148 heads of oncology departments, and 81 (54.7%) questionnaires were completed, with representation from all 17 Spanish autonomous communities. Seventy-seven (95%) of the respondents had EHRs implemented at their hospitals; of them, over 80% considered EHRs to have a positive impact on work organization and clinical practice, and 73% considered that EHRs improve the quality of patient care. In contrast, 27 (35.1%) of these respondents felt that EHRs worsened the physician-patient relationship and conveyed an additional workload (n = 29; 37.6%). Several drawbacks in the implementation of EHRs were identified, including the limited inclusion of information on both outpatients and inpatients, information recorded in free text data fields, and the availability of specific informed consent. Forty-six (56.7%) respondents had patient registries where they recorded information from all patients seen in the department. CONCLUSION: Our study indicates that EHRs are almost universally implemented in the hospitals surveyed and are considered to have a positive impact on work organization and clinical practice. However, EHRs currently have several drawbacks that limit their use for investigational purposes. CLINICAL TRIAL REGISTRATION: Not applicable.

Detection of Salmonella Typhimurium and Listeria monocytogenes biofilm cells exposed to different drying and pre-enrichment times using conventional and rapid methods.

The capacity of real-time PCR (RT-PCR), the VIDAS immunoassay system, and the conventional count method for detecting Salmonella enterica serovar Typhimurium and Listeria monocytogenes biofilm cells was evaluated in this study. After biofilm formation, tests were performed under different drying times (0, 6, 12, 24, and 72 h) and pre-enrichment times (0, 6, 18, and 25 h). The direct epifluorescence microscopic results demonstrated that Salmonella Typhimurium and L. monocytogenes biofilm cells can remain viable for 72 h under drying conditions. Pre-enrichment time and type of medium played an essential role in the detection of both microorganisms after drying. Furthermore, RT-PCR was more sensitive than VIDAS and the conventional method for detecting Salmonella Typhimurium and L. monocytogenes cells at different drying times and without pre-enrichment (0 h), with a detection range between 102 and 107 CFU/mL. TSBYE-T80 used as a pre-enrichment medium was effective for detecting both bacteria and was more effective than Demi Fraser-T80 medium for detecting L. monocytogenes. Therefore, pre-enrichment is recommended to avoid false positives and false negatives due to the presence of dead cells or a very low initial concentration of cells after drying.

New approach for the removal of mature biofilms formed by wild strains of Listeria monocytogenes isolated from food contact surfaces in an Iberian pig processing plant.

One of the main objectives of the food industry is to guarantee food safety by providing innocuous food products. Therefore, this sector must consider all the possible biotic or abiotic contamination routes from the entry of raw materials to the release of the final product. Currently, one important problem in this regard is the presence of biofilms on food contact surfaces which can transmit pathogens such as L. monocytogenes. In industrial conditions biofilms are found in a mature state, so it is essential that when carrying out removal effectiveness studies in vitro the tests are realized with models that produce these structures in a similarly mature state. The main objective of this study was to evaluate the effectiveness of an alternative treatment (i.e. enzymatic detergent that include natural antimicrobial agents) and a conventional treatment (i.e. chlorinated alkaline) for the elimination of mature L. monocytogenes biofilms. The results showed a cell detachment from the formed mature biofilms with an effectivity of between 74.75%-97.73% and 53.94%-94.02% for the enzymatic treatment and the chlorinated alkaline detergent, respectively. On a qualitative level, it was observed that the dispersion in the structure was much higher for the enzymatic treatment than for the chlorinated alkaline, which continued to show obvious structure integrity. All this leads to the conclusion that treatments with an enzymatic detergent have a significantly greater impact on the removal of mature L. monocytogenes biofilms, although a further disinfection process would be needed, enhancing even more the treatment effectivity. This may imply that the industrial approach to addressing this problem should be modified to include new perspectives that are more effective than traditional ones.

Quantification of mature Listeria monocytogenes biofilm cells formed by an in vitro model: A comparison of different methods.

The presence of biofilms in food industrial environments is one of the main causes associated with food product contamination by L. monocytogenes. Biofilm control in the food industry is very relevant to public health and finding reliable and realistic quantification methods is essential. The aim of this study is to compare five L. monocytogenes biofilm quantification methods - conventional plate count, TEMPO, DEM, VIDAS and qPCR - and to examine a biodetector to visually detect biofilms in industrial settings. Results show that depending on the biofilm matrix production, the recovery of cells that conform the biofilm can be low and therefore, if it is an indirect method, microbial counts can be underestimated. At a species level, the methods that did not present significant differences were plate count, TEMPO (P = 0.998), DEM and qPCR (P = 0.508), so correlation studies were performed which established high correlation for plate count and TEMPO, but not for DEM and qPCR. The VIDAS method was adjusted so that it could quantify the biofilms, but the standard curve only allowed counts from 7 Log CFU cm-2. Results also revealed that the different strains of L. monocytogenes possess different biofilm-forming abilities, although it was not possible to correlate the capacity to produce these structures with the distinct serotypes. Last, visually detecting biofilms on stainless steel coupons proved that in industrial environments nowadays they can be rapidly and qualitatively detected so that relevant decisions can immediately be taken.

The Endoscopic Reference Score shows modest accuracy to predict either clinical or histological activity in adult patients with eosinophilic oesophagitis.

BACKGROUND: Conflicting results have been recently reported for the accuracy of the Endoscopic Reference Score (EREFS), an standardised endoscopic classification, to predict the histological activity of eosinophilic oesophagitis (EoE). AIM: To evaluate the accuracy of the EREFS to predict either histological or clinical activity of EoE. METHODS: Prospective multicentre study conducted in eight Spanish centres evaluating adult EoE patients, either naïve or after treatment. Symptoms were evaluated before upper endoscopy through the Dysphagia Symptom Score, whereas researchers scored the EREFS immediately after the endoscopic procedure, unaware of the histological outcome. RESULTS: One hundred and forty-five EoE patients undergoing 240 consecutive endoscopic procedures were included. Exudates (P = 0.03), furrows (P = 0.03) and a composite score of inflammatory signs (exudates, furrows and oedema) (P < 0.001) accurately predicted histological activity. Exudates were the only endoscopic sign showing a good correlation with histological outcome after therapy. Furrows and oedema persisted in 50% and 70% of patients despite histological remission. No endoscopic feature exceeded 70% accuracy to predict histological activity. Likewise, no endoscopic finding could adequately predict dysphagia severity. Crepe paper mucosa, diffuse exudates and severe rings correlated with higher symptom scores. CONCLUSIONS: Endoscopic findings assessed by the Endoscopic Reference Score did not correlate with histological or clinical disease activity in adult EoE patients. Only exudates correlated with peak eosinophil count and histological outcome, whereas furrows and oedema persisted in over half of patients despite histological remission.

Electronic health records and patient registries in medical oncology departments in Spain

Purpose We aimed to evaluate the current situation of electronic health records (EHRs) and patient registries in the oncology departments of hospitals in Spain. Methods This was a cross-sectional study conducted from December 2018 to September 2019. The survey was designed ad hoc by the Outcomes Evaluation and Clinical Practice Section of the Spanish Society of Medical Oncology (SEOM) and was distributed to all head of medical oncology department members of SEOM. Results We invited 148 heads of oncology departments, and 81 (54.7%) questionnaires were completed, with representation from all 17 Spanish autonomous communities. Seventy-seven (95%) of the respondents had EHRs implemented at their hospitals; of them, over 80% considered EHRs to have a positive impact on work organization and clinical practice, and 73% considered that EHRs improve the quality of patient care. In contrast, 27 (35.1%) of these respondents felt that EHRs worsened the physician–patient relationship and conveyed an additional workload (n = 29; 37.6%). Several drawbacks in the implementation of EHRs were identified, including the limited inclusion of information on both outpatients and inpatients, information recorded in free text data fields, and the availability of specific informed consent. Forty-six (56.7%) respondents had patient registries where they recorded information from all patients seen in the department. Conclusion Our study indicates that EHRs are almost universally implemented in the hospitals surveyed and are considered to have a positive impact on work organization and clinical practice. However, EHRs currently have several drawbacks that limit their use for investigational purposes (AU)

Association between Whipple's disease and Giardia lamblia infection.

Whipple' disease is mainly characterized by affecting the digestive system, although it can be a multisystemic process with different clinical symptoms. The bacillus causing the disease has been isolated and cultivated in 2000 and the genome sequence has been recently analyzed in 2003, which means new perspectives for its diagnosis and treatment. Giardiasis is an infestation caused by a protozoo and may cause a malabsorption syndrome or run in a subclinic way. The case of a middle-aged male is described, who after a three-year period of migratory arthralgias, showed weight loss, diarrheas and abdominal pain, being diagnosed of Giardiasis, and after the persistent symptoms and a number of studies, was diagnosed with Whipple disease. Nineteen cases of Giardia-Whipple coinfection have been described in the literature, but the reason of this association has not been found yet. The discussion on whether there is an alteration in the immunitary system which facilitates infections or, the development of an infection lead to the other one, goes on.

Association of hypersensitivity to the nematode Anisakis simplex with HLA class II DRB1*1502-DQB1*0601 haplotype.

Anisakiasis as well as allergic and anaphylactoid reactions to Anisakis simplex antigens are recently identified clinical entities. They are relatively frequent in countries with habitual raw food consumption, often in the form of large amounts of fish and sea food products. In this communication the relationship between HLA class II alleles and the IgE-specific immune response to A. simplex allergen was studied in a defined population in Northern Spain. Individuals with immediate-type Anisakis hypersensitivity and healthy controls were examined for HLA-DRB1, DQB1 and DQA1 alleles by sequence-specific oligonucleotide probe typing. Analysis of the HLA data among patients revealed increased phenotypic frequencies for DRB1*1502 and DRB1*0404 compared to healthy controls (p < 1 x 10(-7) and < 0.01, respectively). Analysis of haplotypic frequencies showed that the DRB1*1502-DQB1*0601 haplotype is significantly higher in patients with Anisakis hypersensitivity in comparison with the control population from the same region (p < 4 x 10(-8)). The data suggest that this haplotype can be considered to be a susceptibility factor for hypersensitivity to A. simplex antigens.

Comparison of staphylococcal co-agglutination with other assays for rapid diagnosis of rotavirus infection in humans, calves and piglets.

Co-agglutination was used for rapid diagnosis of rotavirus infection. Staphylococcus aureus Cowan I sensitized with rabbit antiserum to rotavirus demonstrated the presence of this virus in stools of children, calves and piglets with clinical signs of diarrhea. A total of 415 fecal samples were examined by the co-agglutination test and the results were compared with those of ELISA, electrophoresis in polyacrylamide gel and latex agglutination. The co-agglutination test, when compared to these three methods, presented a correlation which varied between 87.2 and 94.2%.

Incidence of histamine-forming bacteria and histamine content in scombroid fish species from retail markets in the Barcelona area.

Incidence and diversity of histidine decarboxylating bacteria were determined in samples of tunafish, bonito and mackerel purchased at different retail markets. Histamine-forming bacteria occurred in a low proportion and always accounted for less than 0.1% of the total bacterial load in the fish samples studied. Similarly, histamine content in fish samples also was low ( < 25 ppm) and all of them met current histamine standards established by the European Union. Histamine was found in 83.3% of the tested tunafish samples with an average of 8.9 ppm. In contrast, none of mackerel samples and only 2 out of 12 of bonito showed detectable amounts of histamine. Morganella morganii and Klebsiella oxytoca were the most active histamine formers under experimental conditions, and produced on average 2765 and 1415 ppm of histamine, respectively, after incubation at 37 degrees C for 18 h. Some new histamine formers, such as Plesiomonas shigelloides, Enterobacter intermedium, Serratia marcescens, Serratia plymuthica and Serratia fonticola, have been identified. Especially Plesiomonas shigelloides would have an important role within histidine decarboxylating bacteria because it was the sole histamine former isolated that has frequently been associated with the marine aquatic environment. However, only 8-340 ppm of histamine was formed by these strains in laboratory trials.

Protein hydrolysis and proteinase activity during the ripening of salted anchovy (Engraulis encrasicholus l.). A microassay method for determining the protein hydrolysis.

The protein hydrolysis and proteinase activity during the ripening of salted anchovy were studied. A rapid, simple, and inexpensive microassay method for determining the protein hydrolysis by trinitrobenzenesulfonic acid (TNBS) has been developed. A linear relationship was observed between proteolysis determination by the TNBS method and ripening time in the fish muscle and in the brine (r = 0.99). A linear relationship was also observed between the ratio nonprotein nitrogen and total nitrogen (NPN/TN) and ripening time (r = 0.98). Proteolysis by the TNBS method and NPN/TN determination could be considered as objective methods to follow and assess the ripening process of an anchovy. A value of proteolysis by the TNBS method of 240 mM leucine in the fish muscle and/or 200 mM leucine in the brine would indicate the ripening point. The crude enzyme prepared of fish muscle and brine showed that alkaline proteinases dominate.

Myocardial ischemia due to severe amoxicillin allergy.

A patient suffered a myocardial injury as a manifestation of anaphylactic reaction to amoxicillin-clavulanic acid administration. A cardiologic study (ergometry and catheterization) showed no obstructive coronary disease and prick test to amoxicillin was positive. Anaphylaxis may cause myocardial injury and the mechanism is likely to be vasospasm induced by mast cells and basophil mediators.

Monitoring volatile and nonvolatile amines in dried and salted roes of tuna (Thunnus thynnus L.) during manufacture and storage.

Dried and salted roe, obtained from the reproductive organs of female tuna (Thunnus tynnus L.), is a typical fish-based food in the Mediterranean area of Spain. In the present study, we monitored the formation of volatile amines (trimethyamine nitrogen [TMA-N] and total basic volatile nitrogen [TBVN]) and nonvolatile amines (biogenic amines) in dried and salted tuna roe after processing and storage for 8 weeks at 4, 20, and 30 degrees C. The salting and drying process significantly increased the TBVN, cadaverine, tyramine, phenylethylamine, and tryptamine contents, and bacteria with histamine decarboxylase activity were detected both in raw and in dried and salted tuna roes. During storage of tuna roe, TMA-N and TBVN levels increased significantly after the fourth week of storage at 30 degrees C, whereas biogenic amine contents remained more or less constant. However, samples stored at 30 degrees C showed histamine formation after the first week of storage, with a concentration of < 50 ppm. The volatile and nonvolatile amine concentrations in tuna roe were below the consumer safety limit, with the exception of the total biogenic amine level in roe stored at 30 degrees C, which exceeded the European Community's recommended limit (300 ppm). These results indicate that in properly stored tuna roe, histamine formation will not represent a serious health risk to consumers unless the tuna roe has previously been mishandled.

Halotolerant and halophilic histamine-forming bacteria isolated during the ripening of salted anchovies (Engraulis encrasicholus).

This study was performed to investigate halotolerant and halophilic histamine-producing bacteria isolated during the ripening of salted anchovies. Of the isolates obtained during the ripening of anchovies, 1.37% showed histamine-forming activity, most of them (70%) belonging to the Staphylococcus genus. S. epidermidis showed a powerful histamine-forming activity, producing more than 1,000 microg/ml in the presence of 3% and 10% NaCl. Another powerful histamine-producing bacterium isolated during the ripening of salted anchovies was S. capitis. It was able to produce about 400 microg/ml of histamine in 10% NaCl under experimental conditions. Most of these species might be expected to be found as a result of contamination of fish during capture and subsequent unhygienic handling. However, no increase in histamine content was found in any batches through the ripening process. Histamine content always was acceptable in accordance with the maximum allowable levels of histamine fixed by the Spanish and European Union regulations.

An approach to measuring change in muscular tissue contraction.

In this paper the authors consider a histological muscular component from the point of view of systems theory. Of the smooth and striated types of muscular fibres, the latter have a structural component that permits them to contract. Unlike the visceral striated and cardiac fibres, movements in the skeletal striated fibres are predominantly voluntary. An overlapping of certain fibre components--specifically a sliding movement of the filaments of which the fibre is composed--causes a variation in the overall size of the fibre and therefore of the muscle. The distances covered, whether in contraction or lengthening, vary according to the type of fibre from 1.8 to 3.8 microm, but depend only on the capacity of some filaments to slide over other filaments, with filament size itself remaining unchanged. Our objective is to obtain a metric of this system that will permit us to obtain some idea of the degree of variation in these distances. In future projects it will be possible to extend this theory to other involuntary muscular tissue movements (myocardial tissue, e.g.), in an endeavour to develop a mathematical approach to the study of myocardial processes damaged by electrical conduction problems.

Identification of a bisegmented double-stranded RNA virus (picobirnavirus) in calf faeces.

To determine the incidence of rotavirus infection among dairy herds in the State of São Paulo, Brazil, 576 faecal samples obtained from calves aged 1-45 days with and without diarrhoea, reared on 63 dairy cattle farms, were analyzed. Polyacrylamide gel electrophoresis (PAGE) identified 28 samples positive for group A rotavirus, while four samples, two diarrhoeic and two non-diarrhoeic, showed a bisegmented genome with a typical picobirnavirus pattern. Electron microscopy revealed spherical virus particles with a diameter of 37 nm and without a defined surface structure. The present study is the first report of a bisegmented virus identified in cattle in Brazil.