Post-mortem examination and laboratory-based analysis for the diagnosis of bovine tuberculosis among dairy cattle in Ecuador

Veterinary inspection in slaughterhouses allows for the detection of macroscopic lesions reminiscent of bovine tuberculosis, but the presence of Mycobacterium bovis must be confirmed by laboratory methods. This study aimed at comparing the performances of the standard diagnostic tools used to identify M. bovis in tissue specimens sampled from suspicious animals. During a two years period, 1390 cattle were inspected at the Machachi abattoir in the Mejia canton - Ecuador. A total of 33 animals with granulomatous lesions were detected, representing 2.33% (16/687) and 2.42% (17/703) animals examined in 2007 and 2008, respectively. Ninety-four tissue specimens were sampled and screened for the presence of mycobacteria. Acid-fast bacilli were identified in one third of the suspicious cattle (11/33) and suggestive microscopic lesions in 27.3% (9/33) of the samples examined by direct microscopy and histopathology, respectively. Culturing on Stonebrink medium and 16S-rRNA-based polymerase chain reaction (PCR) yielded 36.4% (12/33) and 27.3% (9/33) of positives, respectively. Compared to culture, other diagnostic procedures displayed a lower sensitivity, with 56.5% for PCR, and 43.5% for direct microscopy and histopathology; however, the specificity was higher (94.4% for PCR and microscopy, and 97.2% for histopathology). We conclude that reliable post-mortem laboratory testing either requires the combination of a set of available diagnostic tools or necessitates the development of improved new-generation tools with better sensitivity and specificity characteristics.     

Photocatalytic Removal of the Antibiotic Cefotaxime on TiO<Subscript>2</Subscript> and ZnO Suspensions Under Simulated Sunlight Radiation

This study presents the main results about the removal of the antibiotic cefotaxime (CTX) under simulated sunlight radiation using heterogeneous photocatalysis with titanium dioxide (TiO₂) and zinc oxide (ZnO) in aqueous solutions. The effects of pH and catalyst initial load on pollutant removal were assessed considering the response surface methodology and a central composite circumscribed experimental design, which allowed to determine the optimized conditions that lead to a higher substrate elimination. Experimental results indicated that evaluated parameters have a significant effect on antibiotic removal in both TiO₂ and ZnO suspensions. In addition, the role of photogenerated holes, hydroxyl, and superoxide anion radicals on CTX degradation was evaluated to clarify the reaction mechanism. Finally, analysis of the dissolved organic carbon content in solutions and the antibacterial activity of treated samples showed that photocatalytic treatments were able to reduce a considerable portion of the organic matter present in the systems and its antimicrobial activity.     

Molecular cloning of a Mycoplasma meleagridis-specific antigenic domain endowed with a serodiagnostic potential

A recombinant phage library harbouring Mycoplasma meleagridis (MM) genomic DNA fragments was generated in the bacteriophage lambda gt11 expression vector. The library was screened for expression of MM specific antigens with a polyclonal antiserum that had been preadsorbed with antigens of the most common unrelated avian mycoplasma species. A 49-amino acid antigenic domain unique to MM was isolated, expressed in Escherichia coli, and its serodiagnostic potential was demonstrated. An antiserum raised against this MM-specific antigenic domain recognized a cluster of seven membrane-associated MM proteins with molecular masses ranging from 34 to 75 kDa. Overall, this study resulted in the identification of a potent serodiagnostic tool and revealed the complex antigenic nature of MM.     

Effects of dietary conjugated linoleic acid and high-oleic sunflower oil on performance and egg quality in laying hens

(1) Laying hen performance, yolk fatty acid (FA) concentrations, sensory quality and firmness of eggs were evaluated with respect to the inclusion in the diet of conjugated linoleic acid (CLA) and high-oleic acid sunflower oil (HOSO). (2) Nine diets were arranged factorially, with three concentrations of CIA (0, 1 and 2 g/kg) and HOSO (10, 20 and 30 g/kg). (3) Type of diet did not affect egg production traits. (4) Dietary addition of CLA decreased yolk lipid content and yolk lipid concentrations of monounsaturated FA, C(20:4 n-6) and C(22:6 n-3), but increased those of CLA and saturated FA. (5) Dietary addition of HOSO increased monounsaturated FA concentrations in the yolk lipid but decreased those of CLA and saturated FA. (6) CLA supplementation increased yolk moisture and firmness and impaired the sensory quality of eggs. (7) An interaction between CLA and HOSO addition was found as effects of CLA addition on yolk lipid CLA concentrations and egg quality traits were smaller when the amount of HOSO in the diet increased. (8) Regression equations have been calculated in order to predict yolk CLA and C(18:1), concentration from dietary composition, and yolk firmness from yolk FA composition.     

Does the persistence of sweet chestnut depend on cultural inputs? Regeneration,recruitment, and mortality in <Emphasis Type="Italic">Quercus</Emphasis>- and <Emphasis Type="Italic">Castanea</Emphasis>-dominated forests

Key message

Quercus secondary forests show a gradual transition toward mixed forests, with sweet chestnut ( Castanea sativa ) becoming increasingly abundant in the western Spanish Central System. Additionally, in chestnut-dominated stands, it shows a certain resistance to competitive displacement by Quercus pyrenaica . Our results partially refute the traditional view that C. sativa is unable to recruit in the absence of cultural inputs.

Context

Sweet chestnut, Castanea sativa, is a component of European broadleaf forests and is one of the most managed trees. Due to a reduction in cultural inputs, chestnut-dominated stands tend to be invaded by other species, and it is unclear how chestnut is able to persist in natural mixed forests.

Aims

Our work aimed to identity the main factors that limit the establishment of C. sativa and to analyze the recruitment and mortality processes of C. sativa trees.

Methods

The age, growth ring patterns, regeneration density, and the spatial structure of trees and saplings in 11 plots in the Spanish Central System were analyzed.

Results

Chestnut seedling density increased with C. sativa basal area, but transition toward the sapling stage appeared limited owing to light availability. In Quercus pyrenaica secondary forests, sparse canopies did not constrain chestnut regeneration, and in old chestnut stands, C. sativa showed a certain resistance to competitive displacement. By contrast, mixed young coppices showed a high mortality, most likely due to competition with other vigorous resprouters.

Conclusion

Quercus secondary forests showed a gradual transition toward mixed forests with sweet chestnut becoming increasingly more abundant. In old stands, C. sativa is likely to persist under a gap-phase mode of regeneration. Our results partially refute the traditional view that C. sativa is unable to recruit in the absence of cultural inputs.

     

Mutants in the lipopolysaccharide of Brucella ovis are attenuated and protect against B. ovis infection in mice

Brucella spp. are Gram-negative bacteria that behave as facultative intracellular parasites of a variety of mammals. This genus includes smooth (S) and rough (R) species that carry S and R lipopolysaccharides (LPS), respectively. S-LPS is a virulence factor, and mutants affected in the S-LPS O-polysaccharide (R mutants), core oligosaccharide or both show attenuation. However, B. ovis is naturally R and is virulent in sheep. We studied the role of B. ovis LPS in virulence by mutating the orthologues of wadA, wadB and wadC, three genes known to encode LPS core glycosyltransferases in S brucellae. When mapped with antibodies to outer membrane proteins (Omps) and R-LPS, wadB and wadC mutants displayed defects in LPS structure and outer membrane topology but inactivation of wadA had little or no effect. Consistent with these observations, the wadB and wadC but not the wadA mutants were attenuated in mice. When tested as vaccines, the wadB and wadC mutants protected mice against B. ovis challenge. The results demonstrate that the LPS core is a structure essential for survival in vivo not only of S brucellae but also of a naturally R Brucella pathogenic species, and they confirm our previous hypothesis that the Brucella LPS core is a target for vaccine development. Since vaccine B. melitensis Rev 1 is S and thus interferes in serological testing for S brucellae, wadB mutant represents a candidate vaccine to be evaluated against B. ovis infection of sheep suitable for areas free of B. melitensis.     

Potential application of serological tests on fluids from carcasses: detection of antibodies against Toxoplasma gondii and Sarcoptes scabiei in red foxes (Vulpes vulpes)

Background

Serological surveys for disease investigation of wild animal populations require obtaining blood samples for analysis, which has logistic, ethic and economic difficulties. Applying serological test to fluids collected from dead animals is an alternative. The aim of this study was to assess if antibodies could be detected in two types of fluids collected from 56 carcasses of red foxes (Vulpes vulpes): pleural fluid and lung extract.

Findings

In 22 (39%) foxes antibodies against Sarcoptes scabiei were detected in both fluid types by ELISA and Western blot. In 46 (82%) foxes, antibodies against Toxoplasma gondii were detected in pleural fluid and in 41 (73%) in lung extract applying a Toxo-screen test (DAT). Antibodies were still detectable in the same fluids kept at room temperature for 28 days, although in fewer foxes (16 and 14 foxes tested for T. gondii in lung extract and pleural fluid respectively; and 1 and 4 tested for S. scabiei in lung extract and pleural fluid respectively.

Conclusions

These results indicate the potential utility of using fluids from carcasses for antibody screening of wild animals at the population level.