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Leptospirosis is a bacterial zoonosis in which dogs can act as a reservoir for human infection. The annual vaccination of dogs can prevent leptospirosis caused by serovars included in the vaccine. To date, all available vaccines in Germany include only the serovars Icterohaemorrhagiae and Canicola, the most commonly found serovars prior to the introduction of the leptospirosis vaccines. Yet, the involvement of additional serovars in the clinical presentation of leptospirosis in dogs has been described. The objective of this sero-epidemiological study was to examine the different Leptospira serovars currently circulating in a population of dogs suspicious for leptospirosis from Berlin. In 329 dogs presenting at the Small Animal Clinic in Berlin, the predominant serogroup was Australis (24%), followed by Grippotyphosa (20%) and Pomona (9%). A total of 18% of the dogs were diagnosed with clinical leptospirosis; here the most prevalent serogroups were also Australis (28%), Grippotyphosa (18%), and Pomona (14%). The serovar prevalence data presented here confirm that a change of pattern of infecting Leptospira serovars in dogs has taken place in Berlin. This data corresponds to further sero-epidemiological studies from other regions in Germany. To ensure human and canine health, available vaccines should be adapted to include the most important circulating serovars.