Limited performance of MALDI-TOF for identification of fish Aeromonas isolates at species level.

The aim of this study was to evaluate the usefulness of the MALDI-TOF MS to identify 151 isolates of Aeromonas obtained mostly from diseased fish. MALDI-TOF MS correctly identified all isolates to the genus level but important differences in the percentage of isolates correctly identified depending on the species were observed. Considering exclusively the first identification option, Aeromonas bestiarum, Aeromonas hydrophila, Aeromonas salmonicida, Aeromonas veronii and Aeromonas sobria were the best identified with results >95%. However, considering the first and second identification options, the only species that showed values >90% was A. hydrophila. Overall, when the database was supplemented with 14 new spectra, the number of accurate identifications increased (41% vs. 55%) and the number of inconclusive identifications decreased (45% vs. 29%), but great differences in the success of species-level identifications were found. Species-distinctive mass peaks were identified only for A. hydrophila and A. bestiarum (5003 and 7360 m/z in 95.5% and 94.1% of their isolates, respectively). This work demonstrates the utility of MALDI-TOF MS for Aeromonas identification to the genus level, but there is no consistency for the accurate identification of some of the most prevalent species implicated in fish disease.

Development and evaluation of an IS711-based loop mediated isothermal amplification method (LAMP) for detection of Brucella spp. on clinical samples.

DNA-based methods have emerged as an additional tool for Brucella infection-confirmation at a herd level. However, their implementation may require the use of specialized equipment. In this context the recently developed loop-mediated isothermal amplification (LAMP) technique may constitute an additional and cost-effective tool for rapid and specific DNA detection, especially in low income areas. In the present study the usefulness of a newly developed LAMP assay aiming at the multicopy-IS711 sequence was assessed on a variety of clinical samples (n=81 from abortions and ewes; cattle, n=3; swine, n=4) that were analyzed in parallel using real-time PCR and bacteriology. Although overall sensitivities obtained with the three methods were comparable (p>0.05), our results highlighted the complementarity between bacteriology and molecular-based methods for increased sensitivity. Significant differences (p<0.05) were observed with all techniques depending on the nature of the sample. Our results demonstrate the potential of the IS711-LAMP technique for direct Brucella detection.

Epidemiological factors associated with the exposure of cattle to Coxiella burnetii in the Madrid region of Spain.

Domestic ruminants are considered to be the major source of Coxiella burnetii, the causative agent of Q fever. Even though Q fever is considered to be present worldwide, its distribution in many areas and countries remains unknown. Here, a serological assay was used to estimate the seroprevalence of C. burnetii in cattle in the Madrid region of Spain, to assess its spatial distribution, and to identify risk factors associated with positive results. Ten animals from each of 110 herds (n=1100) were randomly selected and analyzed using an ELISA test. In addition, epidemiological information, at both the herd and individual level, was collected. Variables for which an association with test results was detected in a bivariate analysis were included as predictors (main effects) in a multivariable logistic regression model. Herd and individual seroprevalences were 30% (95% CI=22.2-39.1) and 6.76% (95% CI=5.42-8.41), respectively, and a strong spatial dependence was identified at the first neighbour level using the Cuzick-Edwards test. Production type (dairy >beef >bullfighting) and age of animals (old vs. young) were the only variables significantly associated (P<0.05) with positive serological results at the herd and individual levels, respectively. These results indicate that cattle are exposed to C. burnetii in the Madrid region The high herd seroprevalence found in dairy herds (75%) indicates a higher risk of infection (probably for management reasons) whereas no C. burnetii positive bullfighting herds were identified.