Effect of a quadrivalent vaccine against respiratory virus on the incidence of respiratory disease in weaned beef calves

We investigated the effect of vaccination of male beef calves (mean age+/-S.D.: 158+/-31 days) against bovine herpes virus (BHV-1 or IBR virus), bovine respiratory syncitial virus (BRSV), bovine viral diarrhea (BVD) virus and para-influenza (PI(3)) virus on the incidence of respiratory disease during the first forty days after weaning and entering a feed-lot in Portugal. In May 2003, Mertolenga, Preta and mixed-breed calves from 10 different beef herds, were systematically assigned (by order of entrance in a chute) to two treatment groups, before moving to a common feed-lot. One hundred and twenty five male calves were vaccinated with a quadrivalent vaccine (Rispoval 4) and revaccinated after 21-27 days while 148 herdmates were injected with saline (0.9% NaCl) on the same occasions. The incidence and severity of clinical cases of "bovine respiratory disease" (BRD) were evaluated every day during the first 40 days after entering the feed-lot. Morbidity (3% vs. 14%) and mortality (0% vs. 4%) due to BRD were significantly lower in the vaccinated group. Ten days after revaccination, the calves were treated with an antimicrobial - ending the study - after an outbreak of BRD caused a high incidence of disease in the non-vaccinated group. In conclusion, our results showed that Rispoval 4, a quadrivalent vaccine against respiratory viruses, under field conditions, reduces morbidity and mortality due to BRD in beef calves after weaning.     

Evaluation of cytokine mRNA expression in bronchoalveolar lavage cells from horses with inflammatory airway disease

Inflammatory airway disease (IAD) is a common disorder of performance horses and is associated with poor performance and accumulation of mucus and inflammatory cells in lower airway secretions. Horses with IAD frequently have increased relative counts of neutrophils in bronchoalveolar lavage fluid (BALF); less commonly relative counts of eosinophils and/or mast cells may be increased. The aetiopathogenesis of IAD is unknown and may involve innate and/or acquired immune responses to various factors including respirable dust constituents, micro-organisms, noxious gases and unconditioned air. The molecular pathways and role of the immune system in the pathogenesis of IAD remain poorly defined and it is unknown whether polarised T cell responses occur in the disease, as have been reported to occur in equine recurrent airway obstruction and asthma in humans. Elucidating cytokine responses that develop in horses with IAD may allow a greater understanding of the possible aetiopathological pathway(s) involved and could contribute to development of novel treatments. We compared the mRNA expression of tumour necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), interleukin (IL)-1β, IL-2, IL-4, IL-8, IL-13, IL-17 and IL-23 in cell pellets extracted from BALF of horses with IAD (n=21) and horses free of respiratory tract disease (n=17). Horses with IAD had significantly increased levels of TNF-α, IL-1β and IL-23 mRNA; no significant differences in the other cytokine mRNAs were detected. The results of this study indicate that IAD of horses is associated with increased mRNA expression of pro-inflammatory cytokines in BALF cells, which may reflect stimulation of the innate immune responses to inhaled antigens. There was no evidence of a polarised T-cell cytokine response suggesting hypersensitivity responses may not be involved in the aetiopathogenesis of IAD.     

Molecular cloning and characterization of sea bass (Dicentrarchus labrax L.) CD8alpha

In this work, the gene and cDNA of the sea bass CD8alpha have been isolated and characterized. The coding sequence has an ORF of 666 bp. It retains the Ig motif that interacts with MHC and the two cysteines responsible for an intra-chain disulfide bridge. The hinge region contains the two essential cysteines involved in dimerization. The transmembrane region is well conserved in all analysed sequences. Similar to other teleosts, the cytoplasmic region lacks the consensus p56(lck) motif common in higher vertebrates. Analysis of the expression pattern using RT-PCR shows the highest expression in the thymus. Like in the human gene, the sea bass CD8alpha genomic structure is organized into six exons, which roughly correspond to separate functional domains of the protein. Southern blotting shows that CD8alpha exists as a single copy gene. Together, these results support the concept that the basic structure of CD8alpha has been maintained through evolution.     

First Portuguese isolate of Neospora caninum from an aborted fetus from a dairy herd with endemic neosporosis

Neospora caninum was isolated from the brain of an aborted 4-month-old fetus from a dairy cow herd with endemic neosporosis in Porto, Portugal. The fetal brain homogenate was inoculated interperitoneally first into outbred Swiss Webster mice given dexamethasone and then the peritoneal exudates from these mice was co-inoculated with mouse sarcoma cells in the peritoneal cavity of mice given dexamethasone. N. caninum tachyzoites were seen in peritoneal exudate of the second passage. Tachyzoites from the peritoneal exudate reacted positively with anti-N. caninum antibodies and not with anti-Toxoplasma gondii antibodies and contained N. caninum specific DNA. This Portuguese isolate of N. caninum has been successfully maintained in cell culture. The dam of the aborted fetus had an antibody titer of 1:10240 in the Neospora agglutination test (NAT). Antibodies to N. caninum were found in 76 of 106 cows from this herd in titers of 1:40 in 31, 1:80 in 22, > or =1:160 or more in 23 in the Neospora agglutination test. This is the first isolation of a viable N. caninum-like parasite from any host in Portugal.     

Chronic cork oak decline and water status: new insights

Chronic decline and Sudden death are two syndromes of cork oak (Quercus suber) dieback. Mortality is associated with water stress, but underlying physiological mechanisms are poorly understood. Here, we investigated the physiological performance of declining trees during the summer drought. Leaf water potential, gas-exchange, fluorescence of photosystem II and leaf and root starch concentration were compared in healthy (asymptomatic) and declining trees. Low annual cork increment in declining trees indicated tree decline for several years. All trees showed similar water status in spring. In summer, declining trees showed lower predawn leaf water potential (?2.0 vs. ?0.8 MPa), but unexpectedly higher midday leaf water potential than healthy trees (?2.8 vs. ?3.3 MPa). The higher midday water potential was linked to by means of strongly reduced stomatal conductance and, consequently, transpiration. This study is pioneer showing that declining trees had high midday water potential. A tendency for lower sap flow driving force (the difference between predawn and midday water potential) in declining trees was also associated with reduced photosynthesis, suggesting that chronic dieback may be associated with low carbon uptake. However, starch in roots and leaves was very low and not correlated to the health status of trees. Declining trees showed lower water-use efficiency and non-photochemical quenching in summer, indicating less resistance to drought. Contrarily to chronic decline, one tree that underwent sudden death presented predawn leaf water potential below the cavitation threshold.     

Identification of resistance to rust (Uromyces appendiculatus) and powdery mildew (Erysiphe diffusa) in Portuguese common bean germplasm

Portugal has a diverse common bean germplasm, which is still grown in farmers' fields. In this work, we searched for resistance to rust (Uromyces appendiculatus) and powdery mildew (Erysiphe diffusa) in a representative collection of the Portuguese common bean germplasm. Despite many accessions depicting intermediate levels of resistance when compared to the susceptible check, 24 and 13 accessions showed low levels of infection, in spite of a compatible interaction (disease severity (DS) values lower than 5% and infection type (IT) of 4), to rust and powdery mildew, respectively, indicative of partial resistance. Moreover, a resistant reaction was observed in 11 accessions when inoculated with powdery mildew (IT = 0–1) and in additional 11 accessions (one in common) when inoculated with rust (IT = 0–2). The levels of resistance found in this report anticipate great potential of the Portuguese national germplasm, recently reported as genetically closer to the Andean common bean gene pool, for disease resistance breeding of this important crop.     

Crop and stress coefficients in rainfed and deficit irrigation vineyards using sap flow techniques

Projected climate changes and expansion of viticulture to drier regions justify the installation and management of deficit irrigation (DI) strategies. Contradictory results on the effect of DI on crops may be ascribed to the incorrect application of these techniques. The lack of discrimination between basal crop (K _cb) and stress coefficients (K _s) can be an obstacle to proper irrigation management. A sap flow (SF) technique associated with microlysimeters and eddy covariance (EC) methods was applied to five commercial vineyards, aiming to discriminate those coefficients, during the driest period of the vegetative cycle. A comparative analysis of the coefficients, in relation to measured vegetation parameters (for K _cb) and plant water status (for K _s) is presented. K _cb, ranging from about 0.35 to 0.75, was highly correlated with leaf area index at stand level. K _s, which decreased till 0.2 in the most stressed vineyard, was well correlated to plant water status (K _s function), represented by predawn leaf water potential. K _s functions for the different experiments exhibited falling slopes with decreasing water status, with variable trends depending on the rates of maximal crop transpiration (T _m). These experimental results show that specific parameters for K _s functions, necessary to estimate water use and irrigation depths, in order to control the stress levels in DI scheduling, are also dependent on T _m.     

Influence of wine pectic polysaccharides on the interactions between condensed tannins and salivary proteins

Alpha-amylase, a major human salivary protein, and IB8c, a representative of the proline-rich proteins, were obtained by isolation from saliva and by solid-phase synthesis, respectively. The interactions between these proteins and condensed tannins isolated from grape seeds were studied at different protein and tannin concentrations by measuring their aggregation. Pectic polysaccharides were isolated from wine, and their effect on protein tannin aggregation was assessed. The results presented in this study showed that the most acidic fractions of arabinogalactan proteins have the ability to inhibit the formation of aggregates between the grape seed tannins and the two different salivary proteins. Rhamnogalacturonan II has the same ability toward alpha-amylase but not IB8c under the conditions of the present study. Polysaccharides show effects at concentrations at which they are present in wine, which could mean an influence in wine astringency. The interaction between condensed tannins and alpha-amylase is differently affected by ionic strength when compared with IB8c.     

Inhibition of trypsin by condensed tannins and wine

Phenolic compounds are abundant vegetable secondary metabolites in the human diet. The ability of procyanidin oligomers and wine polyphenols to inhibit trypsin activity was studied using a versatile and reliable in vitro method. The hydrolysis of the chromogenic substrate N-benzoyl-d,l-arginine-p-nitroanilide (BApNA) by trypsin was followed by spectrophotometry in the presence and absence of condensed tannins and wine. A clear relationship between the degree of polymerization of procyanidins and enzymatic inhibition was observed. Trypsin activity inhibition was also detected in several types of wine. In general, the inhibition increased with the concentration of phenolic compounds in wines. These results may be relevant when considering these compounds as antinutritional factors, thereby contributing to a reduced absorption of nutrients.     

A case study of <Emphasis Type="Italic">Eucalyptus globulus</Emphasis> fingerprinting for breeding

• Introduction  

Tree genetic improvement programs usually lack, in general, pedigree information. Since molecular markers can be used to estimate the level of genetic similarity between individuals, we genotyped a sample of a Portuguese Eucalyptus globulus breeding population—a reference population of 125 individuals—with 16 microsatellites (SSR).