Canine vector-borne pathogens are found in many regions and represent a challenge for human and animal health. Shelter dogs are found to be an important health and ecological factor in urban areas in Colombia. Therefore, the aim of the present research was to determine the seroprevalence of canine vector-borne pathogens in shelter dogs from the Metropolitan Area of Bucaramanga, Santander, Colombia. A total of 278 shelter dogs were tested with using a IDEXX SNAP® 4Dx Plus®. From each dog, the following data were collected: municipality, age, gender, and hair length. Overall, 81.3% (226/278) of all tested dogs were positive for one or more of the vector-borne pathogens. The highest seroprevalence observed was Ehrlichia sp. 79.9% (222/278), followed by Anaplasma sp. 36% (100/278), Dirofilaria immitis 6.5% (18/278) and Borrelia burgdorferi 2.2% (6/278). The present research demonstrated the high circulation of vector-borne pathogens among shelter dogs in the Metropolitan area of Bucaramanga, Colombia.