Leptospira spp. serological testing
The presence of serum anti-Leptospira spp. agglutinins was determined by the Microscopic Agglutination Test (MAT). A panel of 24 strains of live Leptospira including the serovars Andamana, Australis, Autumnalis, Bataviae, Bratislava, Butembo, Canicola, Castellonis, Copenhageni, Cynopteri, Grippotyphosa, Hardjo (strain hardjoprajtino), Hebdomadis, Icterohaemorrhagiae, Javanica, Panama, Pomona, Pyrogenes, Sentot, Shermani, Tarassovi, Whitcombi, and Wolffi was used, and a saprophytic serovar Patoc of L. biflexa was used as an internal control. Endpoint titers were determined using two-fold dilutions until the last well showing 50% agglutination was recorded. The final titer was determined by the reciprocal of the positive highest dilution. The cutoff for a positive agglutination reaction was defined as a titer ≥100. For each positive sample, the most probable infecting serogroup was the one presenting the highest titers.