Eosinophilic granulomatosis with polyangiitis: sequential use of mepolizumab following rituximab for inadequate asthma control despite vasculitis remission.

We report the case of a 54-year-old woman with antineutrophilic cytoplasmic antibody-negative eosinophilic granulomatosis with polyangiitis presenting with mononeuritis multiplex, intestinal hemorrhage, cardiomyopathy, fever, and worsening asthma symptoms. She was initially treated with steroids and cyclophosphamide but eventually required rituximab to control a vasculitis flare. However, her asthmatic symptoms did not improve, despite attaining vasculitis remission. Symptoms abated only after the treatment transition to mepolizumab. After a 1-year interval, there were no further episodes of asthma exacerbation and no requirement for systemic steroid therapy. This report reinforces the use of rituximab for induction and maintenance of remission in patients with eosinophilic granulomatosis with polyangiitis and predominant vasculitic manifestations, whereas mepolizumab demonstrated better control of the persistent eosinophilic manifestations, ensuing sustained remission and improved quality of life.

Hepatitis E Virus in Sylvatic and Captive Wild Boar from Portugal.

Hepatitis E virus (HEV) is a zoonotic agent today considered a major Public Health issue in industrialized countries. HEV strains belonging to zoonotic genotype 3 are widely present in swine, being today considered important reservoirs for human disease. Unlike in swine, only scarce data are available on the circulation of HEV in wild boar. This study describes the detection and molecular characterization of HEV in livers from sylvatic wild boar hunted in Portugal and destined for consumption. Additionally, the detection of HEV in stools of a confined wild boar population also destined for consumption is also described. A total of 80 liver samples collected during the hunting season of 2011/2012 and 40 stools collected in February 2012 from a wild boar breeding farm in Portugal were tested by a nested broad-spectrum RT-PCR assay targeting open reading frame (ORF) 1. Twenty livers (25.0%) and 4 stools (10%) were positive for HEV. Phylogenetic analysis showed that all strains clustered with sequences classified as HEV genotype 3 subgenotype e. To our knowledge, this is the first report documenting the occurrence and molecular analysis of HEV in sylvatic and captive wild boar destined for human consumption in Portugal. This report demonstrates for the first time the circulation of HEV in wildlife reservoirs of Portugal adding knowledge to the epidemiology of HEV in wild boar populations.

Lesões melanocíticas em suínos abatidos para consumo / Melanocytic lesions in pigs slaughtered for consumption

O presente estudo teve como principais objetivos conhecer a taxa de ocorrência de lesões melanocíticas (melanose, melanocitomas e melanomas) em suínos abatidos para consumo e identificar possíveis padrões de distinção e de classificação macroscópica dessas lesões. Para tal, procedeu-se à recolha de lesões melanocíticas em matadouro, durante oito meses, e à sua avaliação macroscópica e microscópica. Os resultados deste estudo demonstraram que as melanoses foram as lesões melanocíticas mais frequentemente encontradas (74,04%) e que, relativamente às lesões tumorais, os melanomas (malignos) foram os mais frequentes (21,15%) comparativamente com os melanocitomas (benignos) (4,81%). Pela análise comparativa das características macroscópicas e microscópicas, verificou-se que, nem sempre, por uma avaliação macroscópica, é possível a distinção entre essas lesões (melanoses, melanocitomas e melanomas). No entanto, identificaram-se, neste estudo, algumas características sugestivas da malignidade, como: o tamanho superior a 2,5cm, a presença de ulceração, a libertação de pigmento negro e a presença de coloração negra do gânglio linfático regional. É, portanto, de extrema importância a observação criteriosa e sistemática dessas lesões, para a avaliação das suas características, uma vez que a decisão sanitária é diferente consoante se trate de uma melanose, de um tumor maligno ou de um tumor benigno.

The main objective of the present study was to know the rate of occurrence of melanocytic lesions (melanosis, melanocytoma and melanoma) in pigs slaughtered for consumption and to identify possible patterns of differentiation and a macroscopic classification of these lesions. To this end, we proceeded to the collection of melanocytic lesions in a slaughterhouse during 8 months, and its macroscopic and microscopic evaluation. The results of this study demonstrated that melanosis were the most often found melanocytic lesions (74.04%) and that for tumors, melanomas (malignant) were the most frequent (21.15%) compared to melanocytomas (benign) (4.81%). By comparative analysis of macroscopic and microscopic characteristics we found that it is not always possible, with a macroscopic evaluation, to distinguish between these lesions (melanosis, melanocytoma and melanoma). However, in this study we identified some features suggestive of malignancy such as: size exceeding 2.5cm, the presence of ulceration, the release of black pigment and the presence of a black staining of the regional lymph node. Therefore, it is extremely important to make a careful and systematic observation of these lesions, with the evaluation of their characteristics, since the sanitary decision is different if they are melanosis, malignant tumors or benign tumors.

Risk factors for Salmonella spp in Portuguese breeding pigs using a multilevel analysis.

Salmonella is the second most frequent cause of foodborne illness in the European Union (EU), so EU enforced legislation to achieve a reduction in Salmonella prevalence in the swine sector. To set the reduction target each country carried out a baseline survey to estimate Salmonella prevalence. The aim of our study was to identify risk factors for the presence of Salmonella in breeding pigs based on the data of the Baseline Study for Salmonella in Breeding Pigs in Portugal. In total, 1670 pen fecal samples from 167 herds were tested by culture and 170 samples tested positive. Along with the collection of the samples a survey was applied to collect information about the herd management and potential risk factors. Multilevel analysis was applied to the data using generalized linear mixed models and a logit link function. The outcome variable was the presence/absence of Salmonella in the pen fecal samples. The first level was assigned to the pen fecal samples and the second level to the herds. The results showed significant associations between Salmonella occurrence and the factors (p<0.05): maternity pens versus mating pens (OR=0.39, 95%CI: 0.24-0.63), feed from external or mixed source versus home source (OR=2.81, 95%CI: 1.19-6.61), more than 10 animals per pen versus 10 animals per pen (OR=2.02, 95%CI: 1.19-3.43), North Region versus Alentejo Region (OR=3.86, 95%CI: 1.08-13.75), rodents control (OR=0.23, 95%CI: 0.090-0.59), more than 90% of boars homebred or no boars versus more than 90% of boars from an external source (OR=0.54, 95%CI: 0.3-0.97), semen from another herd versus semen from insemination centers (OR=4.47, 95%CI: 1.38-14.43) and herds with a size of 170 or more sows (OR=1.82, 95%CI: 1.04-3.19). This study offers very relevant information for both the Portuguese veterinary authorities and the pig farmers currently developing control programmes for Salmonella. This is the first study providing evidence for semen and boars source as risk factors for Salmonella in breeding pigs.

Diagnosis of Mycobacterium avium complex in granulomatous lymphadenitis in slaughtered domestic pigs.

The aim of this study was to compare two diagnostic methods for the detection of Mycobacterium avium complex (MAC) infection in lymph nodes with granulomatous lymphadenitis from slaughtered domestic pigs. Fifty affected lymph nodes were collected from 50 pigs and examined microscopically and by polymerase chain reaction (PCR). Microscopically, granulomatous lesions were observed in 92% of the samples, consisting mostly of central necrosis (78%) with dystrophic calcification (46%) and associated with inflammatory infiltration by epithelioid giant cells, lymphocytes, neutrophils (92%), eosinophils (60%) and Langhans-type cells (70%). In 64% of the lesions, a capsule of connective tissue was found. Acid-fast bacilli were observed in all cases. PCR detected DNA from Mycobacterium spp. in 82% (41/50) of the lymph nodes. MAC was confirmed in 58% (24/41) and M. avium avium/silvaticum subspecies in 39% (16/41). The results of this study suggest that combined histopathology and PCR of lymph nodes are useful in the diagnosis of granulomatous lymphadenitis in slaughtered pigs.

No evidence that wild red deer (Cervus elaphus) on the Iberian Peninsula are a reservoir of Mycobacterium avium subspecies paratuberculosis infection.

The potential role of red deer (Cervus elaphus) as a reservoir of Mycobacterium avium subspecies paratuberculosis (MAP) infection is largely unknown. A total of 332 wild red deer were investigated using post-mortem examination, bacteriology and serology. Only three animals (1.12%) were found to have lesions on histopathological examination and no MAP bacteria were recovered on culture. The results suggest it is unlikely that wild red deer make a significant contribution to the maintenance of MAP infection in the region. The cross-reactivity of the ELISAs used indicates this diagnostic modality is ineffective in the detection of MAP infection in this species. The implications of these results for the control of this important pathogen in both livestock and wildlife are discussed.

Genomic and proteomic evaluation of antibiotic resistance in Salmonella strains.

Using Salmonella strains identical to those present in the gastrointestinal tract of different animals we aim to determine and compare the proteome of two serotypes, Salmonella Typhimurium and Enteritidis recovered from faecal samples of wild boars and wild rabbits, respectively. The presence of genes responsible for antibiotic resistance was detected by PCR. Proteomes of the two distinct serotypes were determined using 2-DE in order to identify proteins associated with antibiotic resistance or virulence. Through 2-DE we obtained a total of 229 spots from both strains. All were suitable for MALDI-TOF/TOF and, in correlation with bioinformatic databases, allowed accurate identification and characterization of proteins. S. Enteritidis recovered from wild rabbits was sensitive to all the antibiotics tested in contrast to S. Typhimurium isolated from wild boars which presented a resistance phenotype to ampicillin, streptomycin and chloramphenicol. Nevertheless, despite the different ratio of proteins observed in each proteome according to their biological function, no significant difference was observed in the involvement of these proteins in pathogenicity. Bearing in mind that serotypes are related to infectious processes in humans and animals, it is important to explore the proteome of new strains which might serve as protein biomarkers for biological activity.

Rapid detection of Salmonella sp. in pork samples using fluorescent in situ hybridization: a comparison with VIDAS®-SLM system and ISO 6579 cultural method / Detecção rápida de Salmonella sp. em amostras de suinos por hibridação in situ fluorescente: comparação com o sistema VIDAS®-SLM e com o método de cultura ISO 6579

This study reports the use of the fluorescent in situ hybridization (FISH) with Sal3 probe for Salmonella detection in swine carcasses inner surface (swab); and in the correspondent samples of ileum, ileocolic, and mandibular lymph nodes; and tonsils, after dilution (1:10) in buffered peptone water and a pre-enrichment step (37(0)C, 18h). In order to evaluate the efficiency of FISH, 235 naturally contaminated samples were simultaneously tested by the cultural method (ISO 6579) and by the Vitek Immuno Diagnostic Assay System (VIDAS®) - Salmonella (SLM) system. The cultural method identified 39 positive samples. From these, VIDAS®- SLM only detected 23. FISH identified 115 positive samples. This difference was highly significant (P<0.001). From positive samples, 32 were also confirmed by the cultural method. The results indicate FISH as a promising tool for rapid Salmonella detection in samples of pork and swine carcasses

Descreve-se a utilização da técnica de hibridação in situ fluorescente (FISH), utilizando a sonda Sal3, para detecção de Salmonella na superfície interna de carcaças de suínos (zaragatoa), em amostras correspondentes de íleo, linfonodos ileocólicos, linfonodos mandibulares e amígdalas, após terem sido diluídas (1:10) e submetidas a uma fase de pré-enriquecimento em água peptonada tamponada (a 37ºC, 18h). Para avaliar a eficácia do método FISH, analisaram-se 235 amostras naturalmente contaminadas, usando o método de cultura ISO 6579 e o sistema Vitek Immuno Diagnostic Assay System (VIDAS®)- Salmonella (SLM), simultaneamente. O método de cultura identificou 39 amostras positivas, das quais o método VIDAS®-SLM detectou apenas 23. O método FISH identificou 115 amostras positivas. A diferença entre os métodos foi altamente significativa (P<0.001). Das amostras positivas, 32 foram confirmadas pelo método de cultura. Os resultados indicam que a FISH constitui uma promissora técnica de detecção rápida de Salmonella em amostras de suínos abatidos para consumo

Estado de frescor e qualidade higiênica do pescado vendido numa cidade do interior de Portugal / Freshness and hygienic quality of fish sold in a Portuguese town

Avaliaram-se o estado de frescor e a qualidade higiênica do pescado disponível em alguns estabelecimentos comerciais de Vila Real, uma cidade do interior de Portugal. Vinte e três amostras foram submetidas a uma análise sensorial e a análises fisico-químicas - determinação do teor de azoto básico volátil total (ABVT), do índice de refração do humor aquoso e do pH. A avaliação da qualidade higiênica do pescado foi feita por meio de diferentes análises microbiológicas na superfície e na profundidade do músculo do pescado. O pescado fresco, considerado por todos os métodos de análise, próprio para consumo, apresentou estado de frescor entre bom e satisfatório. O ABVT foi considerado o método objetivo de eleição, sempre que surgiram dúvidas relativamente ao grau de frescor do pescado. O pescado próprio para consumo apresentou valores que variaram entre os 20,60 e 27,36mgN/100g e perfil microbiológico aceitável, com contagens de microrganismos psicrotróficos entre 5,41 e 6,03 UFC/cm² na superfície e entre 4,79 e 5,28 UFC/cm² no músculo

The freshness and hygienic quality of fresh fish sold in some fish markets of Vila Real, a Portuguese town, were evaluated. Twenty-three samples were examined by sensorial and physico-chemical analyses (total volatil basic nitrogen - TVBN, aqueous humour refraction rate and pH). The determination of hygienic quality of fish was carried out by different microbiological analysis on the surface and deepness of fish muscle. Fish was considered proper for consumption by all the methods used in the evaluation of fish freshness, and was classified between good and satisfactory. TVBN was considered the objective method of election, in case of doubt about fish freshness, and fish proper for consumption presented TVBN values between 20.60 and 27.36mgN/100g. The microbiological profile was considered acceptable with psychrotrophic microorganisms counts between 5.41 and 6.03 CFU/cm² on fish surface and between 4.79 and 5.28 CFU/g in muscle deepness

Occurrence of salmonella in the ileum, ileocolic lymph nodes, tonsils, mandibular lymph nodes and carcasses of pigs slaughtered for consumption.

This study evaluates the occurrence of Salmonella in pork carcasses and in some risk tissues (ileum, ileocolic and mandibular lymph nodes and tonsils), that can be involved in Salmonella contamination during slaughter. Salmonella was identified in 27 (26.7%) pigs and in 13 (12.9%) carcasses. From these positive carcasses, 69.2% presented the same serotype as that identified in the corresponding pig, which emphasize the pigs importance as a source of Salmonella during the slaughter, suggesting that measures should be taken at the level of pig production in order to reduce the slaughtering of Salmonella-positive animals. The highest value of Salmonella occurrence was reached in the ileocolic lymph nodes (18.8%) and in the ileum (13.9%), representing Salmonella potential faecal source during pork processing at the abattoir. In these samples, a high level of Salmonella was observed in the ileocolic lymph nodes in comparison with the ileum. The mandibular lymph nodes (12.9%) also presented a higher occurrence in comparison with the tonsils (9.9%). These results indicate that the lymph nodes analysis could be more sensitive in the detection of Salmonella than the closer drainage tissue. Otherwise, the presence of Salmonella in the lymph nodes indicates lymphatic spread of the organism, which reflects an increased risk of pork contamination. These results also indicate that, in order to achieve a better control of Salmonella contamination during the slaughter process, it is important to consider the improvement of the evisceration practices and the tonsils as well the extraction of mandibular lymph nodes after slaughter.