Faculty of Veterinary Medicine University of Zagreb
Abstract
Sera from 120 apparently healthy dogs, 74 purebred, 46 mixed-bred, 57 females and 63 males in the Zagreb area (Croatia) were examined by enzyme-linked immunoadsorbent assay (ELISA) for antibodies to Borrelia burgdorferi. During triple i/v immunization of dog at 1, 7 and 14 day, on each occasion with 5 ml 1.4×107 cells/ml of B. burgdorferi sensu stricto, strain B31 ATCC 35210, a positive control serum was obtained at day 28 p.i., while negative control serum was obtained prior to immunization. The antigen for ELISA was prepared from the same bacterial strain sonicated on ice. The sonicate was quantified by sodium dodecyl sulphate polyacrilamid gel electrophoresis (SDS PAGE). IgG antibodies to B. burgdorferi were estimated in 6 (5%) samples of dog sera in titre ranging from 1:100 to 1:2000 (Optical density-OD=0.650 + 36.49, n=6). Antibodies to B. burgdorferi were found in two females and four males, two Labrador retrievers and in one German shepherd, a Hungarian viszla, one cocker spaniel, and a German wirehaired pointer at 2.5 to 8 years of age. Of 114 seronegative samples three samples were found to be very close to the margin of optical density that determined seropositive result (OD=0.562). Estimated seroprevalence to B. burgdorferi in dog sera suggested that dogs in the Zagreb area are infected with B. burgdorferi and that the Zagreb area is part of a wider Central European Lyme boreliosis endemic area.Imunoenzimnim testom (ELISA) pretraženo je 120 uzoraka klinički zdravih pasa, 74 čistokrvna, 46 križanih, 57 ženki i 63 mužjaka područja grada Zagreba na prisutnost protutijela za B. burgdorferi. Trokratnom i/v imunizacijom psa 1., 7. i 14. dana s po 5 ml kulture B. burgdorferi sensu stricto soj B 31 ATCC 35210 u količini 1,4 × 107 bakterija/ml dobiven je 28. dan od početka imunizacije pozitivni kontrolni serum, a negativni je uzet prije imunizacije. Od istog soja načinjen je ultrazvučno lizirani antigen za imunoenzimni testž (ELISA) čiji je proteinski sastav određen elektroforezom u poliakrilamidnom gelu uz dodatak natrijevog dodecilsulfata (sodium dodecyl sulphate polyacrilamid gel electrophoresis - SDS PAGE). U 6 (5%) uzoraka seruma pasa ustanovljen je titar IgG protutijela od 1:500 do 1:2000 (ODp=0,650 + 36,49, n=6) u dvije kuje i četiri psa u dobi od 2,5 do 8 god., u dva labradora i po jednog njemačkog ovčara, mađarske vižle, koker španijela i njemačkog oštrodlakog ptičara. Od 114 seruma u kojih apsorpcijska vrijednost nije prelazila graničnu apsorpcijsku vrijednost za pozitivne (OD=0,562) u tri seruma apsorpcijska vrijednost je bila blizu granične apsorpcijske vrijednosti. Ustanovljena IgG protutijela za B. burgdorferi u serumima pasa ukazuju da psi područja grada Zagreba bivaju inficirani bakterijom B. burgdorferi i da je to područje dio šireg srednjeeuropskog endemskog žarišta lajmske borelioze
