Update on the role of innate immune receptors during Brucella abortus infection

The innate immune system constitutes an efficient defense mechanism against invading microbial pathogens. Recent studies have revealed the intracellular signaling cascades involved in the TLR-initiated immune response to Brucella spp. infection. However, there is a piece of the puzzle missing that is the role of non-TLR receptors in innate immunity. The involvement of TLR receptors in brucellosis has been investigated by different research groups. It was demonstrated that TLR2 clearly does not play any role in controlling Brucella abortus infection in vivo, whereas TLR9 has been shown to be required for clearance of this bacterium in infected mice. The participation of adaptor molecules, such as MyD88 and TRIF has also been discussed. Recently, we and others have reported the critical role of MyD88- and not TRIF-mediated signaling in dendritic cell maturation and in vivo resistance during B. abortus infection. However, the relationship between specific Brucella molecules and non-TLR receptors and signal transduction pathways needs to be better understood. It is now clear that the interaction between TLRs and recently identified cytosolic innate immune sensors is crucial for mounting effective immune responses. Finally, this review discusses the mechanisms used by Brucella to escape detection by the host innate immune system.     

A multiplex PCR for the detection of Brucella spp. and Leptospira spp. DNA from aborted bovine fetuses

Bovine brucellosis and leptospirosis are important causes of bovine abortion around the world. Both diseases can be serologically diagnosed, but many factors may cause false positive and negative results. Direct methods based on bacteriological isolation are usually employed, but they are difficult, time consuming and dangerous. Monoplex polymerase chain reaction (PCR) have been successfully described for the detection of Brucella spp. and Leptospira spp. Aiming at improvement in the direct diagnosis, a multiplex PCR (mPCR) for the detection of these agents in aborted bovine fetuses is described. The detection threshold of the mPCR was evaluated in experimentally contaminated bovine clinical samples using a conventional proteinase K/SDS or a boiling-based extraction protocols. The mPCR was applied to two groups of clinical samples: 63 episodes of bovine abortion and eight hamsters experimentally infected with Leptospira interrogans serovar pomona. Adopting microbiological isolation as reference, the test showed a sensitivity of 100% in both groups of clinical samples. Seven samples collected from bovine fetuses were Brucella spp. culture negative but showed positive results in mPCR. Regarding Leptospira spp. detection, similar results were observed in three bovine clinical samples. All hamsters infected with Leptospira were positive in both microbiological culture and mPCR. The boiling extraction protocol showed better results in some clinical samples, probably by the removal of PCR inhibitors by heat treatment. The high sensitivity, simplicity and the possibility of detection of both bacteria in a single tube reaction support the use of the mPCR described in the routine diagnosis.     

Selection for contrasting tocopherol content and profile in Ethiopian mustard

Tocopherols are compounds present in oilseeds with major effect on nutritional and technological properties of seed oils. Brassica spp. mainly contain varying concentrations of α‐tocopherol, with high vitamin E value, and γ‐tocopherol, which contributes to oil stability. The objective of this research was to select for contrasting tocopherol content and profile in Ethiopian mustard (Brassica carinata A. Braun). A collection of 211 F_4:5 lines was evaluated for tocopherol content and profile, and divergent selection was conducted till the S_3:4 generation. Selection resulted in two lines with increased tocopherol content (244 and 243 mg/kg seed as average of four environments), one line with reduced tocopherol content (117 mg/kg), two lines with reduced α/γ‐tocopherol ratio (0.22 and 0.28) and two lines with increased α/γ‐tocopherol ratio (3.74 and 3.34). Control line C‐101 averaged 196 mg/kg with α/γ‐tocopherol ratio of 1.10. Reduced α/γ‐tocopherol ratio was controlled by recessive alleles at a single locus. Seed to plant correlation was high (r = 0.92), which suggests the feasibility of selecting at the single‐seed level.     

COMPUTED TOMOGRAPHIC FEATURES OF BASIHYOID ECTOPIC THYROID CARCINOMA IN DOGS

Eight dogs with a firm, nonpainful swelling in the ventral laryngeal region and with a final diagnosis of ectopic thyroid carcinoma were investigated by Computed Tomography (CT) at six different institutions. Computed Tomography findings were reviewed, focusing on lesion volume, shape, margins, relationship with surrounding structures and adjacent vessels, attenuation characteristics, and presence of metastases. Ectopic thyroid carcinomas were seen as oval‐to‐bilobed masses centered on the basihyoid bone with associated bone lysis, highly vascularized capsules with central poorly contrast enhancing areas. In all cases there was laryngeal wall infiltration, in two dogs invasion of the laryngeal lumen and in one case invasion of the ventral muscular and subcutaneous plane. Metastases were found in retropharyngeal lymph nodes (three cases) and in the lung (two cases). Ectopic thyroid carcinoma should be considered in the differential diagnosis when a mass in the basihyoid region is present. Described CT features may be typical for ectopic thyroid neoplasia and could be used to help decide the therapeutic plan.     

Two conifer species native to Patagonia threatened by Phytophthora austrocedri

Phytophthora austrocedri is a pathogen of Austrocedrus chilensis causing “Mal del Ciprés” (cypress sickness) in Patagonia and killing Juniperus communis in Britain, along with other species in the Cupressaceae. The evident association of the pathogen with members of the Cupressaceae makes it necessary to study the susceptibility of other species related to A. chilensis to prevent the emergence of new diseases. The present study tested the pathogenicity of P. austrocedri to three native and endemic conifers in South America: Fitzroya cupressoides, Pilgerodendron uviferum and Araucaria araucana. The results showed that F. cupressoides and P. uviferum were highly susceptible to P. austrocedri. Since these are endangered tree species, the outcome of this study is crucial for governmental agencies, policymakers and stakeholders.     

Assay of the set of all Sudan azodye (I, II, III, IV, and Para-Red) contaminating agents by liquid chromatography-tandem mass spectrometry and isotope dilution methodology

A high-throughput mass spectrometric method is presented for the simultaneous detection of Sudan I, II, III, IV, and Para-Red azodyes in foodstuff. The method is based on the use of deuterium-labeled internal standards and atmospheric pressure chemical ionization (APCI) in a triple-quadrupole instrument. The gas-phase breakdown pattern of each labeled and unlabeled analogue displays the naphthoic moiety as a common fragment. The search for the parents of this common species (parent ion scans) allows, by flow injection and in a single run, the evaluation of the presence of each polluting species spiked in typical foodstuffs. A detailed assay of each azodye was performed by LC-APCI and isotope dilution method, through the multiple reaction monitoring approach, using deuterium-labeled internal standards. Sudan dyes can be quantified above the threshold of 10 ppb except for Sudan Para-Red, for which the limit of quantification was 20 ppb, likely due to the different ionization efficiency.     

An Approach to Air Pollution Source–Receptor Solution by Angular Distances

This study presents the analysis of two series of concentrations of airborne particulate matter (APM) collected in two exploratory campaigns aimed at elucidating the source–receptor problem (SRP) in the metropolitan area of Buenos Aires. Although several techniques have been previously applied to interpret these measurements, we have almost exclusively used here the method of angular distances among objects (ADO) to discuss its advantages as a tool in understanding environmental questions within the source–receptor framework. We present a simple method of calculating the ADO, explain its chemical interpretation and the information that is possible to get by classifying the angular distances. A comparison among ADO with principal component analysis and Kohonen artificial neural networks is also discussed.     

Identification of mycobacterial infections in wild boars in Northern Sardinia, Italy

During a six-month period a region of Northern Sardinia was monitored to check the presence of mycobacterial infections in wild boars. Forty-eight serum and 229 biopsy samples were collected from different animals and examined by both traditional diagnostic techniques (culture, bacterioscopic and molecular tests) and enzyme-linked immunosorbent assay (ELISA). The latter was used to determine the antibody response against both methylated and nonmethylated Heparin-Binding Haemagglutinin (HBHA) protein. Nine mycobacterial strains were isolated: three M. avium ssp. paratuberculosis (Map), three M. avium, one M. interjectum and two M. scrofulaceum strains. By PCR, only one animal was positive for M. bovis, whereas 10 animals were positive for Map. Out of the 48 sera tested, 19 showed a good humoral response to methylated HBHA and 17 to nonmethylated HBHA. Our data provide new information on the prevalence of mycobacterial infection among wild boars in Northern Sardinia and suggest that a more effective program should be developed to monitor mycobacterial infections in the wild animal population.     

Intraspecific breeding for reduced glucosinolate content in Ethiopian mustard (Brassica carinata A. Braun)

A total of six Ethiopian mustard ( Brassica carinata A. Braun) lines showing reduced levels of total glucosinolate content were developed through two different approaches. In the first case, eight lines with good agronomic performance under southern-Spanish conditions were first evaluated during two years for glucosinolate content, and a pedigree selection was then performed during three further generations. The line N2-142, with an average glucosinolate content of 82 μmoles g-1 seed, was developed from the original line C-49, with an average glucosinolate content of 115 μmoles g-1 seed. In the second case, chemical mutagenesis (EMS 1% v/v) was applied to seeds from the line C- 101, with an average glucosinolate content of 125 μmoles g- 1 seed. Five mutant lines showing an average glucosinolate content between 20 and 30 μmoles g-1 seed lower than the wild line C-101 were isolated in the M_3 or M_4 generation. The reduced glucosinolate content of these mutants was confirmed by developing and analysing the M_5 generation. Previous results in B. juncea suggest that a further reduction of total glucosinolate content might be achieved through genetic recombination between the different lines developed in this work. This revised version was published online in July 2006 with corrections to the Cover Date.     

Fluctuating salinity effect on Sphoeroides annulatus (Jenyns 1842) physiological responses

Euryhaline fish, such as the Bullseye puffer Sphoeroides annulatus (Jenyns 1842), experience sudden salinity changes in their natural environment, which is more common than the exception, so they must adapt to survive and cope with extreme salt conditions. Therefore, Bullseye puffer juveniles were exposed to short‐term stress (39 hr) by fluctuating salinity conditions (41, 35, 29, 23, 17, 11, 5, 11, 17, 23, 29, 35, 41 psu) with a 3‐hr interval between each point at 26 ± 1ºC in a respirometer chamber and acclimation reservoirs. Responses to oxygen consumption rate (OCR: 23–35 mg O₂ h^–1 kg^–1), ammonium excretion rate (AER: 1–1.85 mg NH4⁺ h^?1 kg^?1), oxygen‐nitrogen atomic ratio (O:N 17–30), osmoregulatory pattern (blood osmotic pressure from 342.4 to 332.8 mmol/kg) and changes in expression levels of Na⁺/K⁺‐ATPase in the gills (higher values at higher salinities) were measured. Although some signs of stress were detected below the iso‐osmotic point (11.4 psu), the puffer fish is a strong euryhaline fish that survives under these conditions. Nonetheless, it could recover when salinity returned to the initial acclimation point because Sphoeroides annulatus is able to live in a wide range of environments with wide natural salinity fluctuations; thus, a common practice in aquaculture has been to expose fish to low salinity for several reasons discussed in this study. This capacity reveals its high plasticity to saline adaptation from 41 to 5 psu an up from 5 to 41 psu, all in less than 2 days.