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Infected pork meat is an important source of non-typhoidal human salmonellosis. Understanding of molecular mechanisms involved in disease pathogenesis is important for the development of therapeutic and preventive strategies. Thus, hereby we study the transcriptional profiles along the porcine intestine during infection with Salmonella Typhimurium, as well as post-transcriptional gene modulation by microRNAs (miRNA). Sixteen piglets were orally challenged with S. Typhimurium. Samples from jejunum, ileum and colon, collected 1, 2 and 6 days post infection (dpi) were hybridized to mRNA and miRNA expression microarrays and analyzed. Jejunum showed a reduced transcriptional response indicating mild inflammation only at 2 dpi. In ileum inflammatory genes were overexpressed (e.g., IL-1B, IL-6, IL-8, IL1RAP, TNFα), indicating a strong immune response at all times of infection. Infection also down-regulated genes of the FXR pathway (e.g., NR1H4, FABP6, APOA1, SLC10A2), indicating disruption of the bile acid absorption in ileum. This result was confirmed by decreased high-density lipoprotein cholesterol in serum of infected pigs. Ileal inflammatory gene expression changes peaked at 2 dpi and tended to resolve at 6 dpi. Furthermore, miRNA analysis of ileum at 2 dpi revealed 62 miRNAs potentially regulating target genes involved in this inflammatory process (e.g., miR-374 and miR-451). In colon, genes involved in epithelial adherence, proliferation and cellular reorganization were down-regulated at 2 and 6 dpi. In summary, here we show the transcriptional changes occurring at the intestine at different time points of the infection, which are mainly related to inflammation and disruption of the bile acid metabolism.

Context: The Patient Dignity Inventory (PDI) is an instrument to measure sources of distress related to dignity at the end of life., Objectives: To obtain a Spanish version of the PDI and measure psychometric aspects in patients with advanced cancer., Methods: A back-translation method was used to obtain the Spanish version. Inpatients and outpatients with advanced cancer were included. Patients completed the Spanish versions of the PDI (PDI-s), Edmonton Symptom Assessment System (ESAS), Hospital Anxiety and Depression Scale (HADS), and Functional Assessment of Chronic Illness Therapy-Spiritual Well-Being (FACIT-Sp-12) instruments. The psychometric properties evaluated were internal consistency; concurrent validity between PDI-s/ESAS, PDI-s/HADS, and PDI-s/FACIT-Sp-12; discriminant validity, test-retest reliability, and factor analysis. The usefulness of the instrument also was tested., Results: A Spanish version of the PDI was obtained. One hundred twenty-four patients completed the study. Cronbach's alpha coefficient for the PDI-s was 0.89. The PDI-s significantly correlated with the ESAS (rs = 0.669; P < 0.001), HADS (rs = 0.788; P < 0.001), and FACIT-Sp-12 (rs = -0.442; P = 0.008). The instrument distinguished outpatients from inpatients and between patients with differing Karnofsky Performance Status scores (rs = -0.328; P < 0.001). The test-retest method indicated excellent reproducibility (intraclass correlation coefficient = 0.931). Factor analysis showed three factors accounting for 79.4% of the variance. Factors were labeled psychological and existential distress, physical symptoms and dependency, and social support. Patients had no difficulties in understanding or completing the questionnaire (mean time to complete: 7.2 minutes)., Conclusion: The Spanish version of the PDI showed adequate psychometric properties when tested with advanced cancer patients. This research provides a three-factor alternative in Spanish to the PDI., (Copyright © 2015 American Academy of Hospice and Palliative Medicine. Published by Elsevier Inc. All rights reserved.)

Flumequine is a quinolone derivative used in veterinary medicine to treat enteric infections, mainly those caused by Gram negative bacteria and also some Gram positive. Some recent reports by field practitioners have suggested that its use in swine dysentery outbreaks can minimize the impact of this disease. This study aims to evaluate the in vitro anti-Brachyspira hyodysenteriae activity of flumequine. Forty eight field isolates of the bacterium were evaluated using a microdilution test. The lack of colon bioavailability studies of flumequine in pigs makes it difficult to establish the true efficacy of this antibiotic for swine dysentery control. Nonetheless, the relatively high values of MIC50 (50 μg/mL) and MBC50 (50 μg/mL) obtained suggest poor activity against B. hyodysenteriae. Flumequine activity in swine dysentery outbreaks could be related to its activity against other bacteria, different from B. hyodysenteriae, engaged in swine dysentery pathogenesis., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Introduction: To be able to provide patients with a chronic illness personalised care, it is necessary to know how they live with their disorder. Today, there is no validated scale that assesses this aspect., Aim: To present the pilot study of the preliminary version of the living with a chronic process scale (EC-PC, from the name in Spanish) carried out in patients with Parkinson's disease (PD) in Spain., Patients and Methods: A preliminary version of the scale was produced from a literature review and the participation of experts in chronic processes and in PD. Fifteen patients, in all the different stages of PD, answered the preliminary version of the EC-PC (39 items) and a questionnaire about the EC-PC. The viability/acceptability and preliminary aspects of internal consistency were analysed., Results: No domain showed any floor or ceiling effects, but 43.6% of the items had a ceiling effect. The corrected item-total correlation was satisfactory, except in five items. The internal consistency of the five domains was satisfactory, with alpha indexes of 0.81-0.92 and item homogeneity coefficients of 0.19-0.43. The patients identified three items as ambiguous and difficult to answer. Based on these results, 12 items were eliminated and the final version of the EC-PC was drafted, its content being considered satisfactory following its evaluation by expert professionals., Conclusions: The EC-PC, pending validation, is a viable scale of potential interest in the clinical and community healthcare setting for assessing the ability to live with a chronic process like PD.

Porcine epidemic diarrhea (PED) is an enteric disease in swine caused by an alphacoronavirus. It affects swine of all ages causing acute diarrhoea and can lead to severe dehydration and death in suckling piglets. Being recognized for the first time in Europe and Asia during the seventies and the eighties, respectively, it has remained a relevant cause of diarrhea outbreaks in Asia for years and to the present. It has become a major concern in swine production since 2013 when the virus was detected for first time in the USA and in other American countries causing a high number of pig deaths and significant economic losses. The present review aims at approaching the reader to the state of the art of PED giving answer to some of the most recent questions which have arisen related to this disease.

The monophasic Salmonella variant with the antigenic formula Salmonella 4,[5],12:i:- has emerged in the last decade as one of the main serotypes related to human salmonellosis. In the present study, a collection of 94 isolates of the S. 4,12:i:- and S. 4,5,12:i:- coming from Danish farm animals, swine (86), cattle (7), and poultry (1), with well-defined identification was further typed by polymerase chain reaction serotyping, phage typing, and molecular typing (polymerase chain reaction and multilocus variable-number tandem-repeat analysis [MLVA]). Moreover, the determination of antimicrobial resistance pattern of each isolate was tested. In 68 of the isolates the fljB gene was absent (i.e., they were true monophasic strains), whereas in 26 isolates, the gene was present despite the fact that the isolates did not express it. The results clustered the isolates in three main pulse-types. The predominant cluster was compatible with the previously described pattern STYMXB.0131. All the isolates included in this cluster lacked the fljB gene, and all the isolates except one belonged to phage type DT 193 with the AMP-STR-SMX-TET resistance pattern. MLVA analysis divided the clusters in several MLVA profiles previously reported by other studies. Finally, antimicrobial resistance and multiresistance was frequent, although no resistance was detected in critical compounds: fluoroquinolones and cephalosporins. The present study demonstrates the presence of monophasic Salmonella Typhimurium-like strains in Danish food animal production with well-characterized clones that are described by previous studies, demonstrating the emergence and spread of this serotype in Denmark.

Context: The Memorial Delirium Assessment Scale (MDAS) is a reliable and validated instrument with which to assess delirium. However, MDAS responsiveness has only been investigated in an indirect way. Also, neurobehavioral and global cognitive factors seem to be the MDAS main factor loads., Objectives: The primary objective of this study was to evaluate MDAS responsiveness and analyze individual factors on this scale. The secondary objective was to confirm concurrent validity and reliability of the Spanish version of the MDAS., Methods: The translation-back translation method was used to obtain the Spanish version of the MDAS. Delirium diagnosis was determined by the clinical Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision criteria and with the Confusion Assessment Method. Responsiveness and factor loadings were determined with the Delirium Rating Scale-Revised-98, the Mini-Mental State Examination (MMSE), and the MDAS at baseline (0 hours) and at 72 hours., Results: Variation in the scores of the Delirium Rating Scale-Revised-98 shows a correlation of r = 0.93, with variation in MDAS scores at P < 0.001. Variation in MMSE scores shows a correlation of r = -0.84, with variation in MDAS scores at P = 0.015. Factor I, neurobehavioral (reduced awareness, reduced attention, perceptual disturbance, delusions, altered psychomotor activity, and sleep-wake cycle disturbance), correlated moderately with the MMSE at -0.56. Factor II, global cognitive (disorientation, short-term memory impairment, impaired digit span, and disorganized thinking), correlated strongly with the MMSE at -0.81. Factor II was significantly more reliable than Factor I, rho = 0.7, P = 0.01., Conclusion: The high responsiveness confirms the value of the MDAS for ongoing delirium assessment. Two differentiated factor loadings point to a potential future need for MDAS subscales., (Copyright © 2014 U.S. Cancer Pain Relief Committee. All rights reserved.)

In this study, we explored the transcriptional response and the morphological changes occurring in porcine mesenteric lymph-nodes (MLN) along a time course of 1, 2 and 6 days post infection (dpi) with Salmonella Typhimurium. Additionally, we analysed the expression of some Salmonella effectors in tissue to complete our view of the processes triggered in these organs upon infection. The results indicate that besides dampening apoptosis, swine take advantage of the flagellin and prgJ expression by Salmonella Typhimuriun to induce pyroptosis in MLN, preventing bacterial dissemination. Furthermore, cross-presentation of Salmonella antigens was inferred as a mechanism that results in a rapid clearance of pathogen by cytotoxic T cells. In summary, although the Salmonella Typhimurium strain employed in this study was able to express some of its major virulence effectors in porcine MLN, a combination of early innate and adaptive immunity mechanisms might overcome virulence strategies employed by the pathogen, enabling the host to protect itself against bacterial spread beyond gut-associated lymph-nodes. Interestingly, we deduced that clathrin-mediated endocytosis could contribute to mechanisms of pathogen virulence and/or host defence in MLN of Salmonella infected swine. Taken together, our results are useful for a better understanding of the critical protective mechanisms against Salmonella that occur in porcine MLN to prevent the spread of infection beyond the intestine.

Anseriformes deserve special attention in the epidemiology of Brachyspira spp. because diverse Anseriformes species have been described to act as highly efficient carriers of several Brachyspira spp. that can also infect livestock. The aim of this study was to investigate the prevalence and diversity of Brachyspira spp. in waterfowl that winter in Spain. Brachyspira spp. were isolated from 51 of the 205 faecal samples collected from graylag geese and mallards in the Villafáfila Lagoons Nature Reserve (Northwestern Spain). The Brachyspira species identified through phenotyping, PCR and sequencing of the nox gene were B. pilosicoli (5.9%), B. alvinipulli (11.8%), "B. hampsonii" (19.6%), B. murdochii (23.5%) and B. innocens (39.2%). The most relevant finding of this study is the description of "B. hampsonii" in specimens from birds for the first time. Phylogenetic analysis of the nox gene sequences grouped all of the obtained "B. hampsonii" isolates into a cluster with Brachyspira strains previously identified by others as "B. hampsonii" and separated from other Brachyspira spp. isolates and reference strains. Additionally, this cluster was related to clades that grouped B. murdochii and B. innocens isolates. The identification of "B. hampsonii" was also achieved in 8 of the 10 isolates by sequencing the16S rRNA gene and tlyA gene. Regardless of the species identified, no antimicrobial resistance was observed in any of the enteropathogenic isolates recovered. This is the first description of "B. hampsonii" in European waterfowl, which might represent hosts that serve as natural reservoirs of this Brachyspira species. This finding indicates that this spirochete is not limited to North America, and its presence in wild birds in Europe poses a risk of transmission to livestock.

The objective of this paper is to analyse in further detail the Danish results of the EFSA baseline studies in slaughter pigs and breeding herds, and compare them with the results obtained in (1) the pre-implementation study that was carried out to establish the initial prevalence values in fattening herds as part of the Danish Salmonella control programme, and (2) the study performed four years later in breeding and finishing herds to obtain information about the prevalence in breeding farms and the status of the finishers after the first years of the National Salmonella Control Programme. In the slaughter pigs Salmonella was detected in a 7.4% of 1218 ileocaecal lymph nodes and on 3.2% of 438 carcasses examined. Among the breeding herds examined by floor faecal or swab samples 122 of 298 (40.9%) were positive in at least one of the ten samples collected. The most prevalent serotypes were Salmonella Typhimurium in finishers and Salmonella Derby in breeding herds while the most prevalent phage types of the S. Typhimurium isolates were DT 12 and DT 120. The antimicrobial resistance analysis yielded a 35.2% of the isolates from the slaughter pigs resistant to one or more antimicrobials while 19.3% were resistant to four or more antimicrobials. A significantly higher percentage of resistance to antimicrobials was found in the S. Typhimurium isolates (χ(2)=4.72, p=0.029), where 42.9% presented resistance to one or more compounds. In breeding herds, just S. Typhimurium and S. 4,5],12:i: - isolates were tested. As many as 56.8% of the S. Typhimurium-like strains positive breeding farms had resistant strains, while 27% had multidrug resistant strains. The distribution of the isolates in regions showed that S. Derby is at present the predominant serotype in breeding farms from most of the regions of the country., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Pork is a major cause of foodborne salmonellosis. Consequently, effective measures that could reduce the prevalence of Salmonella at the farm are of interest. In the present study, three field trials were performed to evaluate the effect of strategic administration of organic acids, at concentrations estimated by economic criteria, on the Salmonella prevalence in finishing pigs. Pigs received either a mixture of acids (lactic, formic, propionic, and acetic) added to their drinking water at a concentration of 0.035% (trial A), or a basal diet containing 0.5% potassium-diformate, KH(COOH)₂, (trials B and C), during the last 6-7 weeks of the finishing period. Fecal Salmonella shedding and seroprevalence were monitored in the animals at four time points during the study. Mesenteric lymph nodes and cecal contents were collected from a subset of animals at the slaughterhouse and cultured for Salmonella. At the end of the finishing period in all three trials, the percentage of seropositive pigs was higher in the control group than in the experimental group, regardless of which cutoff value was used in the enzyme-linked immunosorbent assay (ELISA) assay. The frequency of fecal shedding was lower also in the treated pig groups from the trials A and B at the end of the finishing period. The results from the samples collected at the slaughterhouse did not yield differences between groups in trials A and B. However, Salmonella isolation was less frequent in the mesenteric lymph nodes from the experimental pigs in trial C. The seroprevalence reductions, together with some promising results in the reduction of shedding, support the idea that this intervention is a useful strategy to reduce Salmonella prevalence at the farm.

The efficacy of an inactivated S. Typhimurium vaccine administered to pigs at the beginning of the fattening period was evaluated in four clinical trials (trials A, B, C and D). Faecal shedding and the systemic antibody response during fattening, as well as, the cecal contents and mesenteric lymph nodes collected after slaughtering were used to assess the outcome. Salmonella shedders prevalence in the control groups was six times higher than in the treated groups in trials A and D, both herds infected by S. Typhimurium. The risk of positive pens was also four or five times higher for the pens housing control pigs in trials A and C. Lower prevalence of Salmonella was observed in the slaughter samples from the vaccinated pigs in trial D and in the cecal content samples in trial A, when just the S. Typhimurium results were compared. The results suggest the effective homologous protection of the vaccinated pigs; however, the high humoral response elicited in the vaccinated pigs complicates their use in farms under serological surveillance programmes., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

In this report we employed laser-capture microdissection (LCM) coupled to qPCR technology and bioinformatic analysis to characterize, for the first time, the response of Peyer's patches (PP) from orally infected animals to Salmonella typhimurium, in a model of non-typhoidal salmonellosis. Pathogen was highly found in the cytoplasm of phagocytes in PP and differential gene expression analysis indicated an up-regulation of proinflammatory molecules, establishment of a Th1 driven response and triggering of DC and T-cell activity. Furthermore, predictions by bioinformatic analysis pointed to an activation of processes regarding stimulation and maturation of DC, influx of leukocytes in tissue and T lymphocytes priming and differentiation. In short, the approach used in this study proved to be a promising strategy to explore infectious processes. Indeed, it revealed an effective induction of innate and adaptive immune mechanisms in swine PP which appear to be distinct from those observed in mesenteric lymph nodes and closely related to response of gut mucosa., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Swine Dysentery (SD) is a severe mucohaemorhagic enteric disease of pigs caused by Brachyspira hyodysenteriae, which has a large impact on pig production and causes important losses due to mortality and sub-optimal performance. Although B. hyodysenteriae has been traditionally considered a pathogen mainly transmitted by direct contact, through the introduction of subclinically infected animals into a previously uninfected herd, recent findings position B. hyodysenteriae as a potential threat for indirect transmission between farms. This article summarizes the knowledge available on the etiological agent of SD and its virulence traits, and reviews the determinants of SD transmission. The between-herds and within-herd transmission routes are addressed. The factors affecting disease transmission are thoroughly discussed, i.e., environmental survival of the pathogen, husbandry factors (production system, production stage, farm management), role of vectors, diet influence and interaction of the microorganism with gut microbiota. Finally, prophylactic and therapeutic approaches to fight against the disease are briefly described.

Salmonella is one of the major foodborne pathogens worldwide. Pork products are among the main sources of Salmonella infection in humans, and several countries have established Salmonella surveillance and control programs. The role of slaughtering in carcass contamination has been indicated by studies focused on the slaughterhouse environment. In this review, we examine and discuss the information available regarding the influence that farm status, pig transport, and lairage have on the carriage of Salmonella by pigs entering the slaughter line. The evolution of carcass contamination throughout the slaughtering process, the main sources of contamination in the dirty and clean zones of the slaughter line, and previously reported prevalence of Salmonella on carcasses and factors affecting this prevalence also are discussed. The importance of implementing interventions at the slaughter level is discussed briefly. Consistent with the information available, pigs from infected farms and newly acquired or recrudescent infections in pigs at the subsequent stages of transport and lairage are important sources of Salmonella at the slaughtering plant. The continuous introduction of Salmonella into the slaughterhouse and the potential for resident flora constitute a risk for carcass contamination. At the slaughterhouse, some dressing activities can reduce carcass contamination, but others are critical control points that jeopardize carcass hygiene. This information indicates the importance of considering slaughter and previous stages in the pork production chain for controlling Salmonella in swine production.

Infections caused by Salmonella enterica serovar Typhimurium (S. typhimurium) cause important economic problems in the swine industry and threaten the integrity of a safe and healthy food supply. Controlling the prevalence of Salmonella in pig production requires a thorough knowledge of the response processes that occurs in the gut associated immune tissues. To explore the in vivo porcine response to S. typhimurium, MLN samples from four control pigs and twelve infected animals at 1, 2 and 6 days post infection (dpi) were collected to quantify the mRNA expression of gene coding for 42 innate immune-related molecules. In addition, the presence of S. typhimurium in MLN was examined and its effect on tissue micro-anatomy. Higher S. typhimurium loads were observed at 2dpi, triggering an innate immune response, marked by a substantial infiltration of phagocytes and up-regulation of pro-inflammatory genes. Such response resulted in a significant decrease in pathogen burden in MLN at 6dpi, although Salmonella could not be completely eliminated from tissue. Furthermore, our results suggest that in porcine infections, S. typhimurium might interferes with dendritic cell-T cell interactions and this strategy could be involved in the conversion of Salmonella infected pigs to a carrier state., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The objective of this study was to investigate the role of the slaughtering process in Salmonella carcass contamination by typing isolates recovered previously in a double study of the following: (1) a tracking survey from the farm to the slaughterhouse and (2) a survey of the slaughterhouse environment (i.e., lairage area, slaughter line, cutting plant and carcasses). The Salmonella serotypes identified on the carcasses of the 16 tracked batches were frequently linked to lairage, whereas the serotypes detected at the farm, transport or pig-related samples (i.e., caecum content and lymph nodes) were only occasionally detected at the carcass level. Multi-locus variable-number tandem repeats (MLVA) of 77 Salmonella enterica ser. Typhimurium isolates from seven of these batches confirmed the link between the isolates recovered from carcasses and holding pens. Only four of the 16 positive carcasses had profiles previously isolated from lymph nodes or caecal content. In the second part of the study, a total of 131 S. enterica ser. Typhimurium and 74 S. enterica ser. Derby isolates were further characterised by MLVA and Pulsed Field Gel Electrophoresis (PFGE), respectively. The MLVA profiles identified in carcasses varied throughout the working day and were frequently linked to those identified in samples from the slaughter line points collected close in time. PFGE and MLVA profiles identified at lairage were also detected in later processing facilities (i.e., slaughter line and cutting plant) as well as in carcasses. Finally, most of the profiles found at the cutting plants were previously identified in the slaughter line or carcass samples. The results from this study show that Salmonella contamination in pigs entering the slaughterhouse can be attributed to several sources. Typing of isolates by MLVA and PFGE clarified the sources of carcass contamination and improved the accuracy of cross-contamination attributable values. Without obviating the relevant role of infected pigs entering the slaughterhouse, the present study highlights the lairage and slaughtering as important sources of carcass contamination., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Context: The Edmonton Symptom Assessment System (ESAS) is a measure widely used in palliative care for the assessment of symptoms in patients with advanced cancer. The tool has been validated in different languages, including Spanish. A revised version (ESAS-r) was developed by Watanabe et al. in 2010., Objectives: To develop the Spanish version of the ESAS-r and examine its psychometric properties., Methods: Based on the original English version, a group of experts created a Spanish version of the ESAS-r and administered it to a group of advanced cancer patients. Patients completed the ESAS and ESAS-r and were asked for their perceptions of the tool. The psychometric properties of the ESAS-r that were analyzed were equivalence, internal consistency, and discriminant validity., Results: Sixty-six patients from Spain and Guatemala participated in the survey. Patients perceived the ESAS-r to be significantly easier to understand and easier to complete than the ESAS. Significantly, patients preferred the ESAS-r (47%) to the ESAS (15%; P<0.0007). As to reliability, we found good internal consistency (Cronbach's alpha 0.86), and the equivalence of the two versions was between 0.71 and 0.94. The ESAS-r discriminates between inpatients and outpatients (Mann-Whitney U test; P=0.02) and among those with different palliative performance status (Spearman's rho for pain, tiredness, drowsiness, lack of appetite, well-being; P<0.01)., Conclusion: The ESAS-r is a valid instrument with adequate psychometric characteristics. This version is preferred by patients with advanced cancer. The Spanish version of the ESAS-r can, therefore, replace the use of the ESAS., (Copyright © 2013 U.S. Cancer Pain Relief Committee. Published by Elsevier Inc. All rights reserved.)

In this study we employed for the first time an in vivo approach coupled to DIGE-based proteomics to explore the response of porcine mesenteric lymph nodes (MLN) to Salmonella typhimurium infection. MLN samples were collected from four control and twelve infected pigs (at 1, 2 and 6 days post infection) for histological analysis, protein and RNA purification. Afterwards, expressed proteins were screened by differential in gel analysis and data were analyzed by bioinformatic tools to generate interaction networks, and identify enriched signaling pathways and biological annotations. S. typhimurium labeling in tissue and phagocyte infiltration were analyzed by immunohistochemistry and RNA was employed to determine the relative expression of immune-related genes by quantitative RNA analysis. The proteome response of porcine MLN to infection was associated to the induction of processes such as phagocyte infiltration, cytoskeleton remodeling and pyroptosis. Moreover, our results suggest that S. typhimurium antigens are cross-presented via MHC-I in a proteasome-dependent manner in porcine MLN. Since pathogen burden in tissue was noticeably reduced at the end of the time course, we infer that host innate and adaptive immunity act in association in MLN to control S. typhimurium dissemination in swine infections., (Copyright © 2012 Elsevier B.V. All rights reserved.)

The enteropathogen Salmonella Typhimurium is one of the main causes of porcine and human enterocolitis. We have used a 2-DE, MALDI-TOF/TOF-based approach to characterize in vivo proteome changes in porcine ileum mucosa after pathogen interaction. Ileum samples from non-infected and orally infected animals were collected at 2 days post infection and S. Typhimurium presence was confirmed by immunohistochemistry. Fifty one proteins, involved in immune response (acute phase response, inflammation and immune response regulation), apoptosis and pathogen-mediated cell invasion, were identified as being differentially expressed after pathogen challenge. Overall, anti-inflammatory signals and a possible down-regulation of dendritic cell maturation were observed. According to this, we identified the up-regulation of FK506-binding protein 4 (FKBP4), a negative regulator of the transcription factor IRF4 (interferon regulatory factor 4), implicated in Th2 and Th17 response. Transcriptional analysis using RT-qPCR indicated a general trend toward down-regulation of Th2 and Th17 cytokines genes, which would be in agreement with an IRF4 reduced transactivation activity. On the other hand, proteins that could be involved in maturation of Salmonella-containing vacuole and intracellular pathogen survival were up-regulated. Results derived from this study would be valuable to better characterize a possible pathogen led modulation of host responses in vivo., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Background: Swine dysentery (SD) is a widespread diarrhoeal disease of pigs caused by infection of the large intestine with the anaerobic intestinal spirochaete Brachyspira hyodysenteriae. Understanding the dynamics of SD, and hence being able to develop more effective measures to counter its spread, depends on the ability to characterise B. hyodysenteriae variants and trace relationships of epidemic strains., Methodology/principal Findings: A collection of 51 Spanish and 1 Portuguese B. hyodysenteriae isolates was examined using a multilocus sequence typing (MLST) scheme based on the sequences of seven conserved genomic loci. The isolates were allocated to 10 sequence types (STs) in three major groups of descent. Isolates in four of the STs were widely distributed in farms around Spain. One farm was infected with isolates from more than one ST. Sequence data obtained from PubMLST for 111 other B. hyodysenteriae strains from other countries then were included in the analysis. Two of the predominant STs that were found in Spain also were present in other European countries. The 73 STs were arranged in eleven clonal complexes (Cc) containing between 2 and 26 isolates. A population snapshot based on amino acid types (AATs) placed 75% of the isolates from 32 of the 48 AATs into one major cluster. The founder type AAT9 included 22 isolates from 10 STs that were recovered in Spain, Australia, Sweden, Germany, Belgium, the UK, Canada, and the USA., Conclusions/significance: This MLST scheme provided sufficient resolution power to unambiguously characterise B. hyodysenteriae isolates, and can be recommended as a routine typing tool that rapidly enables comparisons of isolates. Using this method it was shown that some of the main genetic lineages of B. hyodysenteriae in Spain also occurred in other countries, providing further evidence for international transmission. Finally, analysis of AATs appeared useful for deducing putative ancestral relationships between strains.

Background: The Edmonton Symptom Assessment System (ESAS) is developed for daily symptom assessment. Validation studies tested a variety of languages and patients. The purpose was to carry out a comprehensive examination of the psychometric properties of the ESAS through validation of the version in Spanish advanced cancer patients., Method: A reverse translation method was used to translate the ESAS. Previous studies find appropriate Spanish terms to explore, with verbal scales, fatigue, depression and anxiety. Psychometric aspects evaluated were reliability, validity, responsiveness and utility., Results: 171 advanced cancer patients participated. Internal consistency with Cronbach's Alpha was 0.75. In test-retest (0-6h), Spearma's correlation was between 0.65 and 0.94. Factor analysis found 3 central domains: 'soft' and 'hard physical' and 'emotional'. Concurrent validity with the Rotterdam Symptom Check List (RSCL) found good correlation in physical symptoms (Kappa until 0.66) but weak correlation in emotional symptoms (Kappa 0.35). Discriminant validity (Spearman) found significant differences (p<0.001) classifying by Karnofsky. ESAS discriminate between inpatients and outpatients (Mann-Whitney, p<0.001). Responsiveness was tested with ESAS at 0-48 h (Wilcoxon test, p<0.05). Average time to complete the instrument was 5.5 min., Conclusion: ESAS is a valid, reliable, responsive and feasible instrument with adequate psychometric properties when tested on Spanish advanced cancer patients., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The antimicrobial susceptibility of clinical isolates of Brachyspira hyodysenteriae in Spain was monitored, and the underlying molecular mechanisms of resistance were investigated. MICs of tylosin, tiamulin, valnemulin, lincomycin, and tylvalosin were determined for 87 B. hyodysenteriae isolates recovered from 2008 to 2009 by broth dilution. Domain V of the 23S rRNA gene and the ribosomal protein L3 gene were sequenced in 20 isolates for which the tiamulin MIC was ≥ 4 μg/ml, presenting decreased susceptibility, and in 18 tiamulin-susceptible isolates (MIC ≤ 0.125 μg/ml), and all isolates were typed by multiple-locus variable-number tandem repeats analysis. A comparison with antimicrobial susceptibility data from 2000 to 2007 showed an increase in pleuromutilin resistance over time, doubling the number of isolates with decreased susceptibility to tiamulin. No alteration in susceptibility was detected for lincomycin, and the MIC of tylosin remained high (MIC(50) > 128 μg/ml). The decreased susceptibility to tylosin and lincomycin can be explained by mutations at position A2058 of the 23S rRNA gene (Escherichia coli numbering). A2058T was the predominant mutation, but A2058G also was found together with a change of the neighboring base pair at positions 2057 to 2611. The role of additional point mutations in the vicinity of the peptidyl transferase center and mutations in the L3 at amino acids 148 and 149 and their possible involvement in antimicrobial susceptibility are considered. An association between G2032A and high levels of tiamulin and lincomycin MICs was found, suggesting an increasing importance of this mutation in antimicrobial resistance of clinical isolates of B. hyodysenteriae.

The aims of this study were to investigate the prevalence of colonization with intestinal spirochaetes in dogs, and to assess their association with diarrhoea. To achieve this, faecal samples from 311 dogs were obtained between November 2008 and April 2009 and cultured for Brachyspira species. A total of 41 Brachyspira spp. isolates were recovered, and these were classified into species according to their biochemical properties, and results of a B. pilosicoli species-specific PCR, and partial amplification of the nox gene with sequencing of the product. An overall Brachyspira spp. prevalence of 13.2% (41/311) was obtained. The prevalence of Brachyspira pilosicoli faecal shedding was 4.8% (15/311) while "Brachyspira canis" was identified in 8.0% (25/311) of the sampled dogs. One dog shed an isolate tentatively identified as B. intermedia. A statistically significant association between the shedding of B. pilosicoli and the presence of diarrhoea in dogs was demonstrated (P<0.001). Risk factors for shedding of Brachyspira spp. were investigated. Using the odds ratio, the risk of B. pilosicoli shedding was five times higher among dogs up to 1 year of age as compared with adult dogs (older than 1 year). These findings may have practical implications in the public and animal health fields., (Copyright © 2010 Elsevier B.V. All rights reserved.)

The spirochete Brachyspira hyodysenteriae is the causative agent of swine dysentery, a severe colonic infection of pigs that has a considerable economic impact in many swine-producing countries. In spite of its importance, knowledge about the global epidemiology and population structure of B. hyodysenteriae is limited. Progress in this area has been hampered by the lack of a low-cost, portable, and discriminatory method for strain typing. The aim of the current study was to develop and test a multiple-locus variable-number tandem-repeat analysis (MLVA) method that could be used in basic veterinary diagnostic microbiology laboratories equipped with PCR technology or in more advanced laboratories with access to capillary electrophoresis. Based on eight loci, and when performed on isolates from different farms in different countries, as well as type and reference strains, the MLVA technique developed was highly discriminatory (Hunter and Gaston discriminatory index, 0.938 [95% confidence interval, 0.9175 to 0.9584]) while retaining a high phylogenetic value. Using the technique, the species was shown to be diverse (44 MLVA types from 172 isolates and strains), although isolates were stable in herds over time. The population structure appeared to be clonal. The finding of B. hyodysenteriae MLVA type 3 in piggeries in three European countries, as well as other, related, strains in different countries, suggests that spreading of the pathogen via carrier pigs is likely. MLVA overcame drawbacks associated with previous typing techniques for B. hyodysenteriae and was a powerful method for epidemiologic and population structure studies on this important pathogenic spirochete.

Salmonella enterica serovar Typhimurium causes enteric disease and compromises food safety. In pigs, the molecular response of the intestine to S. typhimurium has been traditionally characterized by in vitro models that do not reflect the actual immunological competence of the intestinal mucosa. In this work, we performed an oral S. typhimurium infection study to obtain insight into the in vitro response in three different sections (jejunum, ileum and colon) of the porcine intestine. For this, samples from one-month-old infected piglets were collected during a time course comprising 1, 2, and 6 days post inoculation to evaluate the intestinal response by quantifying the mRNA expression of gene coding for 28 innate immune system molecules using quantitative real-time PCR assays. In addition, samples from non-infected control animals were also employed to establish differences in the steady state gene expression between intestinal sections. The panel of quantified molecules included an assortment of cytokines, chemokines, pattern-recognition receptors, intracellular signaling molecules, transcription factors and antimicrobial molecules. Changes in gene expression occurred in the three different parts of the intestine and during the course of the S. typhimurium infection. Moreover, the high variation observed in expression patterns of genes coding for inflammatory mediators could indicate that each intestinal section responds differently to the infection. Thus, on the contrary to findings in the jejunum and colon, a down-regulation and lack of induction of some proinflammatory cytokine transcripts was observed in the ileum. Nevertheless, all chemoattractant cytokines assayed were up-regulated in the ileum and jejunum whereas only interleukin-8 and MIP-1alpha mRNA were over expressed in the colon. In conclusion, our results reveal regional differences in gene expression profiles along the porcine intestinal gut as well as regional differences in the inflammatory response to S. typhimurium infection. Taken together, these data should provide a basis for a complete understanding of the porcine intestinal response to bacterial infection., (INRA, EDP Sciences, 2010)

Herd-level risk factors for faecal shedding of Salmonella enterica were investigated in a cross-sectional study on Spanish finishing units. For this purpose, 10 faecal samples were collected from 10 different pens containing pigs close to market weight in a total of 232 fattening units. The total sample size was proportionally distributed according to the fattener census in each of the regions and provinces of the country in order to ensure a sample representative of the entire swine population. All samples were individually examined by culture of 25g of faecal material. Data regarding characteristics and management of each fattening unit were collected by means of a questionnaire. Logistic regression was used to detect relationships between the detection of faecal shedding of S. enterica and potential herd-level risk factors. The feeding of pelleted feed was associated with an increased risk of culture-positive faecal samples (OR=2.28; 95% CI: 1.22, 4.26). The odds of a farm being Salmonella positive were associated with its size. Fattening units that slaughtered more than 3500 pigs per year had a higher risk for Salmonella faecal shedding (OR=1.78; 95% CI: 0.96, 3.31). Interventions at these two points should be considered when designing or managing growing pig facilities to reduce Salmonella faecal shedding by fatteners.

The effect of hemoglobin (Hb) and lipid concentration, pH, temperature, and different antioxidants on heme-mediated lipid oxidation of liposomes from marine phospholipids was studied. The rate of lipid oxidation was measured by consumption of dissolved oxygen. Heme-mediated lipid oxidation at different Hb and lipid concentrations was modeled by Michaelis-Menten kinetics. The maximum rate (V(max)) for the reaction with cod and bovine Hb as a pro-oxidant was 66.2 +/- 3.4 and 56.6 +/- 3.4 microM/min, respectively. The Michaelis-Menten constant (K(m)) for the reaction with cod and bovine Hb was 0.67 +/- 0.09 and 1.2 +/- 0.2 microM, respectively. V(max) for the relationship between the oxygen uptake rate and lipid concentration was 43.2 +/- 1.5 microM/min, while the K(m) was 0.93 +/- 0.14 mg/mL. The effect of the temperature followed Arrhenius kinetics, and there was no significant difference in activation energy between cod and bovine Hb. The rate of lipid oxidation induced by bovine Hb was highest around pH 6. Ethylenediaminetetraacetic acid (EDTA) had no significant effect on heme-mediated lipid oxidation, but alpha-tocopherol and astaxanthin worked well as antioxidants. Kinetic differences were found between iron and Hb as pro-oxidants, and the efficacy of the antioxidants depended upon the pro-oxidant in the system.

On screening tools for emotional distress, the terms "depression" and "anxiety" are commonly used for patients with advanced cancer. However, these terms could have negative connotations in Spanish such that cultural and unexpected differences in perception may invalidate or skew the results of the screening if the best terms are not chosen. The goal of this study was to determine the best expression that can be used to explore anxiety and depression in Spanish. A prospective study of 100 Spanish-speaking patients was performed. Spanish patients with cancer completed the Hospital Anxiety and Depression Scale (HADS) and six Verbal Numerical Scales (VNS) exploring the level of anxiety using the terms ansioso (anxious), nervioso (nervous), or intranquilo (uneasy/disquiet), and the level of depression using the terms deprimido (depressed), desanimado (discouraged), or triste (sad). The correlation, sensitivity, and specificity for all the VNS and HADS (8 and 11 cutoff points) were analyzed. The correlation (Spearman rho) between HADS and the anxiety VNS was r = 0.557 using "anxious"; r = 0.603 using "nervous"; and r = 0.594 using "uneasy." The correlation for the depression VNS was r = 0.662 using "depression"; r = 0.759 using "discouraged" and r = 0.596 using "sad"; alpha < 0.001 was used in all VNS. A cutoff point of 4 of 10 for any term used to explore anxiety achieved the best levels for sensitivity (0.80) and specificity (0.70). The term "discouraged" with a cutoff point of 4 of 10 shows a sensitivity of 0.89, a specificity of 0.84, as well as a predictive positive value of 0.77 and a negative value of 0.93. In Spanish, the term desanimado seems to be more suitable in screening for depression. Alternate terms could be used to explore anxiety in Spanish. Exploring depression with simple questions in Spanish achieves greater accuracy than the same approach to exploring anxiety.

Introduction: Fatigue is one of the most frequent symptoms in patients with cancer. No adequate term in Spanish has been defined to describe the English concept of fatigue., Aim: To identify the most suitable Spanish words that define the concept of fatigue and to check psychometric characteristics., Method: Consensus with professional experts on Spanish words that best suit the English concept of fatigue. A prospective study on oncologic patients was also undertaken, which included an evaluation of the intensity of fatigue through visual numeric scales (VNS) where the words had been previously selected. The fatigue subscale of the Functional Assessment of Cancer Therapy-Fatigue (FACT-F) questionnaire was taken as a reference., Results: The experts highlighted the words cansancio, agotamiento, and debilidad (tiredness, exhaustion, and weakness) as the terms that best defined the concept of fatigue. In the psychometric assessment study, 100 patients were included, of which 61 (61%) presented diagnostic values for cancer-related fatigue in the FACT-F fatigue subscale (score 34/52 or lower). The VNS for the chosen terms obtained a high correlation with the FACT-F fatigue subscale results: cansancio (tiredness) r = -0.71, agotamiento (exhaustion) r = -0.74, debilidad (weakness) r = -0.74, with no statistical differences between them. For the detection of fatigue by means of the VNS, tiredness (cutoff point > or =4/10) gave sensitivity (S) 0.90 and specificity (E) 0.72; exhaustion (cutoff point > or =3/10) S 0.95 and E 0.90 and weakness (cutoff point > or =4/10) S 0.92 and E 0.72. The ROC curve was 0.88 for tiredness, 0.94 for exhaustion, and 0.92 for weakness, with no significant difference between the areas mentioned. The terms cansancio, agotamiento, and debilidad (tiredness, exhaustion, and weakness) are suitable for defining the English concept of fatigue in Spanish, and should be the preferred option for inclusion in evaluation tools.

It is not clear if 18FDG-PET can be useful for detection of inflammation in low to moderate carotid stenosis. We studied 15 patients scheduled for endarterectomy with contralateral carotids with less than 50% stenosis. 18-FDG-PET was performed prior to CEA and 3 months following surgery. FDG-uptake values were calculated based on maximum standardized uptake value (SUV) and corresponding uptake ratios. We confirmed by CD68 macrophage staining that FDG accumulation corresponds to active inflammation (R=0.8 p less than 0.005). We found significant correlation between the FDG-uptake in the carotids scheduled for CEA and contralateral carotids with low to moderate stenosis (R=0.9 p less than 0.001). The FDG uptake ratio in the contralateral arteries remained stable on the follow-up imaging (1.15+/-0.2 vs. 1.14+/-0.1, R=0.7 p=0.006). We did not find correlation between FDG uptake and symptomatic or asymptomatic patients, degree of carotid stenosis and vascular risk factors. This is a prospective, preliminary in vivo study demonstrating that low to moderate carotid atherosclerosis can be detected using 18-FDG-PET imaging and highlights the truly systemic nature of atherosclerosis.

The European mink, Mustela lutreola, has suffered a dramatic decline in Europe during the 20th century and is one of the most endangered carnivores in the world. The subpopulation of European mink from Navarra, Spain, estimated to number approximately 420, represents approximately two thirds of the total number of mink in Spain. Aleutian Disease Virus (ADV) is a parvovirus with a high degree of variability that can infect a broad range of mustelid hosts. The pathogenesis of this virus in small carnivores is variable and can be influenced by both host factors (e.g., species, American mink genotype, and immune status) and viral strain. A cross-sectional study was conducted during the pre-reproductive period of February-March 2004 and 2005 and the postreproductive period of September-December 2004. Mink were intensively trapped along seven rivers that were representative of the European mink habitat in Navarra. Antibody counter immunoelectrophoresis against ADV was performed on 84 European mink blood samples. All the samples were negative. Protein electrophoresis was performed on 93 plasma samples. Nine of those samples (9.6%) had gamma globulin levels exceeding 20% of the total plasma protein. Complete necropsies were performed on 23 cadavers of European mink collected in the area between 2000 and 2005. Seventeen of the mink (74%) had traumatic and hemorrhagic lesions compatible with vehicular impact injuries. Although there were no histopathologic lesions associated with ADV, this study documents the first description of a naturally occurring canine distemper virus infection in a European mink. In addition, pulmonary adiaspiromycosis in three European mink from Spain was reported.

Hypoxia, angiogenesis and inflammation leads to plaque progression and remodelling and may significantly contribute towards plaque rupture and subsequent cerebrovascular events. Our aim was to study, markers of hypoxia and inflammation previously identified by microarray analysis, in atherosclerotic carotid arteries with low to moderate stenosis. We hoped to describe different cellular populations expressing the studied markers. The location of selected inflammatory molecules obtained as vascular transplants from organ donors were analysed by immunohistochemistry with monoclonal and polyclonal antibodies. Paraffin-embedded sections were cut and probed with antibodies recognizing active B and T-lymphocytes (CD30), hypoxia-inducible factor-1alpha, endoglin (CD105), Interleukin-6 and C-reactive protein. We observed a notable overexpression of HIF-1alpha in inflammatory and hypoxic areas of carotid arteries in all types of lesions from type II-V taken from the patients with carotid stenosis less than 50%. This suggests that HIF-1alpha may have a putative role in atherosclerosis progression and angiogenesis. Dynamic changes in the non-occluding plaques may explain some of the clinical events in patients with low to moderate carotid stenosis.

Background and Purpose: There is growing evidence suggesting that C-reactive protein (CRP) is an effecter molecule able to induce and promote atherothrombosis. The presence of CRP in atherosclerotic plaques may reflect local production or infiltration from circulating CRP increased in general inflammatory responses. Our aim was to analyze the presence of CRP in human advanced carotid artery plaques with differential anatomo-pathological characteristics and to assess local expression of CRP and other proinflammatory genes in these lesions., Methods: Human carotid artery specimens from 38 patients undergoing scheduled endarterectomy were classified into 3 groups: ulcerated (noncomplicated) (UNC, n=19), fibrous (F, n=12) and ulcerated (complicated/hemorrhagic) plaques (UC, n=7). The presence of CRP was evaluated by immunohistochemistry, and plasma samples were screened for circulating high-sensitivity C-reactive protein. TaqMan Low-density Arrays were used for study of genes related to inflammation (CRP, interleukin-6, macrophage colony-stimulating factor-1, monocyte chemotactic protein-1, cyclooxygenase-2)., Results: CRP mRNA levels were predominantly detected in UNC-high risk plaques but not in UC (P=0.001). UNC also exhibit the highest expression levels of other genes involved in the inflammatory responses: cyclooxygenase-2 (P<0.005 versus F and versus UC), IL-6 (P<0.005 versus F and versus UC) and monocyte chemoattractant protein-1 (P<0.01 versus F and versus UC). Plaque CRP mRNA levels correlated with immunohistochemical findings but were independent of plasma high-sensitivity CRP. In UNC plaques endothelial cells and inflammatory cells were strongly positive for CRP around areas of newly formed microvessels., Conclusions: In human high-risk carotid artery plaques (UNC) CRP expression reflects an active proinflammatory stage. Local synthesis of CRP could be involved in plaque neovascularization and increased risk of hemorrhagic transformation.

The genetic diversity of the Derby serotype of Salmonella enterica in Spain was examined by pulsed-field gel electrophoresis (PFGE). Out of 24 identified PFGE profiles, a major clone was detected in 19% of strains from humans, 52% from food, and 62% from swine. This clone (clone 1) was isolated from pork products, suggesting swine as its source.

Objective: Osteomyelitis due to Candida species is an unusual but recognized entity. However, with the increasing occurrence of factors predisposing to candidemia and invasive candidiasis, candidal osteomyelitis is being diagnosed more frequently. An unusual case of Candida albicans osteomyelitis is reported here, along with a review of the published literature on previously reported cases of this disease., Methods: Report of the case and literature review., Results: In this case, Candida albicans was isolated from the talus; however, the diagnosis was made after several cultures were performed. Only one other case of Candida osteomyelitis located in foot bones was found in the review., Conclusions: It is thought that this is the first case reported in Venezuela, and only the second in international literature.