Lisbon, Portugal
Lisbon, Portugal

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Martins A.P.,New University of Lisbon | Lopes P.A.,CIISA | Martins S.V.,CIISA | Madeira A.,New University of Lisbon | And 6 more authors.
Biochemical and Biophysical Research Communications | Year: 2010

Conjugated linoleic acid (CLA) is a dietary fatty acid frequently used as a body fat reducing agent whose effects upon cell membranes and cellular function remain unknown. Obese Zucker rats were fed atherogenic diets containing saturated fats of vegetable or animal origin with or without 1% CLA, as a mixture of cis(c)9,. trans(t)11 and t10,. c12 isomers. Plasma membrane vesicles obtained from visceral adipose tissue were used to assess the effectiveness of dietary fat and CLA membrane incorporation and its outcome on fluidity and permeability to water and glycerol. A significant decrease in adipose membrane fluidity was correlated with the changes observed in permeability, which seem to be caused by the incorporation of the t10,. c12 CLA isomer into membrane phospholipids. These results indicate that CLA supplementation in obese Zucker rats fed saturated and cholesterol rich diets reduces the fluidity and permeability of adipose membranes, therefore not supporting CLA as a body fat reducing agent through membrane fluidification in obese fat consumers. © 2010 Elsevier Inc.


Coelho J.,CIISA | Martins C.,CIISA | Ferreira F.,CIISA | Leitao A.,University of Lisbon
Virology | Year: 2015

Topoisomerases modulate the topological state of DNA during processes, such as replication and transcription, that cause overwinding and/or underwinding of the DNA. African swine fever virus (ASFV) is a nucleo-cytoplasmic double-stranded DNA virus shown to contain an OFR (P1192R) with homology to type II topoisomerases. Here we observed that pP1192R is highly conserved among ASFV isolates but dissimilar from other viral, prokaryotic or eukaryotic type II topoisomerases. In both ASFV/Ba71V-infected Vero cells and ASFV/L60-infected pig macrophages we detected pP1192R at intermediate and late phases of infection, cytoplasmically localized and accumulating in the viral factories. Finally, we used a Saccharomyces cerevisiae temperature-sensitive strain in order to demonstrate, through complementation and in vitro decatenation assays, the functionality of P1192R, which we further confirmed by mutating its predicted catalytic residue. Overall, this work strengthens the idea that P1192R constitutes a target for studying, and possibly controlling, ASFV transcription and replication. © 2014 Elsevier Inc.


Baptista F.M.,Copenhagen University | Alban L.,Danish Agriculture and Food Council | Nielsen L.R.,Copenhagen University | Domingos I.,CIISA | And 2 more authors.
Zoonoses and Public Health | Year: 2010

Salmonella surveillance-and-control programs in pigs are highly resource demanding, so alternative cost-effective approaches are desirable. The aim of this study was to develop and evaluate a tool for predicting the Salmonella test status in pig herds based on herd information collected from 108 industrial farrow-to-finish pig herds in Portugal. A questionnaire including known risk factors for Salmonella was used. A factor analysis model was developed to identify relevant factors that were then tested for association with Salmonella status. Three factors were identified and labelled: general biosecurity (factor 1), herd size (factor 2) and sanitary gap implementation (factor 3). Based on the loadings in factor 1 and factor 3, herds were classified according to their biosecurity practices. In total, 59% of the herds had a good level of biosecurity (interpreted as a loading below zero in factor 1) and 37% of the farms had good biosecurity and implemented sanitary gap (loading below zero in factor 1 and loading above zero in factor 3). This implied that they, among other things, implemented preventive measures for visitors and workers entering the herd, controlled biological vectors, had hygiene procedures in place, water quality assessment, and sanitary gap in the fattening and growing sections. In total, 50 herds were tested for Salmonella. Logistic regression analysis showed that factor 1 was significantly associated with Salmonella test status (P = 0.04). Herds with poor biosecurity had a higher probability of testing Salmonella positive compared with herds with good biosecurity. This study shows the potential for using herd information to classify herds according to their Salmonella status in the absence of good testing options. The method might be used as a potentially cost-effective tool for future development of risk-based approaches to surveillance, targeting interventions to high-risk herds or differentiating sampling strategies in herds with different levels of infection. © 2010 Blackwell Verlag GmbH.


Alho A.M.,CIISA | Schnyder M.,University of Zürich | Schaper R.,Bayer Animal Health GmbH | Meireles J.,CIISA | And 3 more authors.
Parasitology Research | Year: 2016

Angiostrongylus vasorum is a nematode that lives in the pulmonary arteries and right cardiac ventricle of domestic dogs and wild canids. It is increasingly being reported in several European countries and North America. This parasite induces inflammatory verminous pneumonia, causing severe respiratory disease in dogs. In some instances, coagulopathies, neurological signs and even death may occur. Scant data are available regarding the occurrence of A. vasorum in Portugal. Therefore, sera of 906 shelter dogs from North to South mainland Portugal were collected. ELISAs to detect A. vasorum circulating antigen and specific antibodies against this parasite were performed. A total of six dogs [0.66 %, 95 % confidence intervals (CI) 0.24–1.43] were positive for both A. vasorum antigen and antibody detection, indicating an active infection, and 12 dogs (1.32 %, CI 0.68–2.30) were A. vasorum antibody-positive only. Regions with antigen- and antibody-positive animals overlapped and were distributed over nearly all sampled areas in the country. This is the first large-scale ELISA-based serological survey for A. vasorum in dogs from Portugal. The endemic occurrence of A. vasorum in dogs from different geographical areas of Portugal is therefore confirmed. © 2016 The Author(s)


PubMed | University of Zürich, CIISA, New University of Lisbon and Bayer Animal Health GmbH
Type: Journal Article | Journal: Parasitology research | Year: 2016

Angiostrongylus vasorum is a nematode that lives in the pulmonary arteries and right cardiac ventricle of domestic dogs and wild canids. It is increasingly being reported in several European countries and North America. This parasite induces inflammatory verminous pneumonia, causing severe respiratory disease in dogs. In some instances, coagulopathies, neurological signs and even death may occur. Scant data are available regarding the occurrence of A. vasorum in Portugal. Therefore, sera of 906 shelter dogs from North to South mainland Portugal were collected. ELISAs to detect A. vasorum circulating antigen and specific antibodies against this parasite were performed. A total of six dogs [0.66%, 95% confidence intervals (CI) 0.24-1.43] were positive for both A. vasorum antigen and antibody detection, indicating an active infection, and 12 dogs (1.32%, CI 0.68-2.30) were A. vasorum antibody-positive only. Regions with antigen- and antibody-positive animals overlapped and were distributed over nearly all sampled areas in the country. This is the first large-scale ELISA-based serological survey for A. vasorum in dogs from Portugal. The endemic occurrence of A. vasorum in dogs from different geographical areas of Portugal is therefore confirmed.


Matos B.,CIISA | Colella V.,University of Bari | Alho A.M.,CIISA | Otranto D.,University of Bari | And 2 more authors.
Helminthologia (Poland) | Year: 2016

Here we report the case of a diagnostic and treatment challenge in a four-month old dog from England, presented with one-month history of unproductive cough. Antigen blood test for Angiostrongylus vasorum infection was negative. Thoracic radiographs revealed a generalised bronchointerstitial pattern and bronchoscopy showed moving nematodes in the mucus of the bronchial wall. Additionally, Baermann technique revealed a high burden of larvae per gram of faeces. Morphological and molecular analyses confirmed that they were first stage larvae of Crenosoma vulpis. The infection was firstly treated with a spot-on solution containing 10% imidacloprid + 2.5% moxidectin, but the dog was still positive after 13 days. Therefore, a seven-day course of fenbendazole was prescribed. This represents one of the youngest dogs ever reported naturally infected by C. vulpis. The scant number of reported cases of crenosomosis has led practitioners to consider it as a rare parasitic disease in dogs, delaying a correct and targeted on-time diagnosis. Further studies are needed to perceive the real prevalence of this lungworm and to understand if it is a rare parasite or just rarely diagnosed. © 2016 Institute of Parasitology, SAS, Košice 2016.


PubMed | Hospital Veterinario das Laranjeiras, Hospital Veterinario Inemvet, CIISA and University of Bari
Type: Journal Article | Journal: Parasitology international | Year: 2016

Onchocerca lupi (Spirurida, Onchocercidae) is an emerging vector-borne helminth that causes nodular lesions associated with acute or chronic ocular disease in dogs and cats. Since its first description in dogs in 1991, this zoonotic filarioid has been increasingly reported in Europe and the United States. An 8-year-old outdoor mixed-breed female dog from the Algarve (southern Portugal) was presented with a history of severe dyspnoea. Cervical and thoracic radiographs revealed a slight reduction in the diameter of the cervical trachea and a moderate increase in radiopacity of the laryngeal soft tissue. An exploratory laryngoscopy was performed, revealing filiform worms associated with stenosis of the thyroid cartilage and a purulent necrotic tissue in the larynx lumen. A single sessile nodule, protruding from the dorsal wall of the laryngeal lumen caused a severe reduction of the glottis and tracheal diameter. Fragments of the worms were morphologically and molecularly identified as O. lupi. Histological examination of the nodule showed a granulomatous reaction with sections of coiled gravid female nematodes. Following laryngoscopy, a tracheostomy tube was inserted to relieve dyspnoea and ivermectin (300 g/kg, once a week, for 8 weeks) combined with prednisolone was prescribed. The dog showed a complete recovery. Although O. lupi has been isolated in human patients from the spinal cord, this is the first report of an aberrant migration of O. lupi in a dog. The veterinary medical community should pay attention to aberrant location of O. lupi and consider onchocercosis as a differential diagnosis for airway obstruction in dogs.


Couto M.,CIISA | Santos A.S.,Escola Universitaria Vasco da Gama | Laborda J.,Associacao para o Estudo e Protecao do Gado Asinino | Novoa M.,Associacao para o Estudo e Protecao do Gado Asinino | And 2 more authors.
Livestock Science | Year: 2016

The Miranda donkey (Equus asinus) has an important role in the maintenance of rural landscape and traditions in inland of Portugal. Breeding and keeping of these animals rely on mountain pasture areas, known for their characteristic botanical diversity. However, their grazing behaviour in these areas has not been described and this knowledge is important to set the best management practices that promote an efficient use of these particular feed resources that could be related with parasitic level of these animals. In this way, this study aimed to assess feeding behaviour, diet selection, and relate the total phenolic and tannin content of ingested plants with the parasite load in a group of Miranda donkeys under free ranging system in natural mountain pastures. For this purpose, eight adult non-pregnant Jennies from the Miranda donkey breed were managed under continuous grazing on a 1.6 ha natural mountain pasture from late May to late July. Body condition score (BCS) and grazing behaviour of the Jennies was assessed in two different occasions. Additionally, the level of gastrointestinal nematode infections of the herd was assessed throughout the study. Results showed that animals spent 75.6% of total observation time in search/prehension activities corresponding to an average of 16 h/day. Jennies showed a preference for herbaceous species, although they were able to incorporate in their diet up to 30% of shrubs, suggesting that they can be used as biological tool for controlling and reducing shrub encroachment in these mountain areas. Parasite level increased from 0% positive samples in June to a maximum of 25% in July. The increase of EPG may be related with the observed decrease in the shrub consumption throughout the observation period. Having in account the presence of phenolic and condensed tannins in shrubs, their potential as natural anti-parasitic compounds and the possibility of taking part in donkey diet, these preliminary results can be of the utmost importance regarding an integrated approach of Miranda donkey parasite control. © 2016 Elsevier B.V.


Ferreira-Dias G.,CIISA | Costa A.S.,CIISA | Mateus L.,CIISA | Korzekwa A.J.,Institute of Animal Reproduction and Food Research | And 7 more authors.
Domestic Animal Endocrinology | Year: 2011

Cytokines and nitric oxide (NO) are potential mediators of luteal development and maintenance, angiogenesis, and blood flow. The aim of this study was to evaluate (i) the localization and protein expression of endothelial and inducible nitric oxide synthases (eNOS and iNOS) in equine corpora lutea (CL) throughout the luteal phase and (ii) the effect of a nitric oxide donor (spermine NONOate, NONOate) on the production of progesterone (P4) and prostaglandin (PG) E2 and factor(s) that stimulate endothelial cell proliferation using equine luteal explants. Luteal tissue was classified as corpora hemorrhagica (CH; n = 5), midluteal phase CL (mid-CL; n = 5) or late luteal phase CL (late CL; n = 5). Both eNOS and iNOS were localized in large luteal cells and endothelial cells throughout the luteal phase. The expression of eNOS was the lowest in mid-CL (P < 0.05) and the highest in late CL (P < 0.05). However, no change was found for iNOS expression. Luteal explants were cultured with no hormone added or with NONOate (10-5 M), tumor necrosis factor-α (TNFα; 10 ng/mL; positive control), or equine LH (100 ng/mL; positive control). Conditioned media by luteal tissues were assayed for P4 and PGE2 and for their ability to stimulate proliferation of bovine aortic endothelial cells (BAEC). All treatments stimulated release of P4 in CH, but not in mid-CL. TNFα and NONOate treatments also increased PGE2 levels and BAEC proliferation in CH (P < 0.05). However, in mid-CL, no changes were observed, regardless of the treatments used. These data suggest that NO and TNFα stimulate equine CH secretory functions and the production of angiogenic factor(s). Furthermore, in mares, NO may play a role in CL growth during early luteal development, when vascular development is more intense. © 2011 Elsevier Inc.


PubMed | CIISA
Type: Journal Article | Journal: Domestic animal endocrinology | Year: 2010

Cytokines and nitric oxide (NO) are potential mediators of luteal development and maintenance, angiogenesis, and blood flow. The aim of this study was to evaluate (i) the localization and protein expression of endothelial and inducible nitric oxide synthases (eNOS and iNOS) in equine corpora lutea (CL) throughout the luteal phase and (ii) the effect of a nitric oxide donor (spermine NONOate, NONOate) on the production of progesterone (P4) and prostaglandin (PG) E(2) and factor(s) that stimulate endothelial cell proliferation using equine luteal explants. Luteal tissue was classified as corpora hemorrhagica (CH; n = 5), midluteal phase CL (mid-CL; n = 5) or late luteal phase CL (late CL; n = 5). Both eNOS and iNOS were localized in large luteal cells and endothelial cells throughout the luteal phase. The expression of eNOS was the lowest in mid-CL (P < 0.05) and the highest in late CL (P < 0.05). However, no change was found for iNOS expression. Luteal explants were cultured with no hormone added or with NONOate (10(-5) M), tumor necrosis factor- (TNF; 10 ng/mL; positive control), or equine LH (100 ng/mL; positive control). Conditioned media by luteal tissues were assayed for P4 and PGE(2) and for their ability to stimulate proliferation of bovine aortic endothelial cells (BAEC). All treatments stimulated release of P4 in CH, but not in mid-CL. TNF and NONOate treatments also increased PGE(2) levels and BAEC proliferation in CH (P < 0.05). However, in mid-CL, no changes were observed, regardless of the treatments used. These data suggest that NO and TNF stimulate equine CH secretory functions and the production of angiogenic factor(s). Furthermore, in mares, NO may play a role in CL growth during early luteal development, when vascular development is more intense.

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