MOLECULAR DETECTION AND SEROPREVALENCE OF MYCOPLASMAS IN CLINICALY HEALTHY WORKING DOGS
Authors: Scholten Sara Suhadolc , Tozon Nataša , Koprivec Saša , Felda Kaja , Florjančič Mateja , Benčina Dušan , Slavec Brigita 
Published: July 2018

In this study seroprevalence and prevalence of mycoplasmas in clinically healthy dogs were studied. Thirty-four working dogs of various breeds, gender and age were included in this research. Among them, 27 were working dogs from Slovene armed forces and 7 were working sheepdogs. We used dot-immunobinding assay (DIBA) as a serological test for the detection of specific antibodies to Mycoplasma cynos, Mycoplasma canis and Mycoplasma molare and consensus PCR for detection of genes for 16S rRNA or 16S/23S IGS region of mycoplasmas. Specific antibodies against at least one of the canine mycoplasmas were detected in 94.1% dogs. Of them 23.5% samples showed positive reaction only to M. cynos, 20.6% were positive only to M. canis and none of the samples were positive only to M. molare. Altogether 47.0% of samples were positive to M. cynos and M. canis whereas only one dog (2.9%) had specific antibodies to all three mycoplasmas tested. The presence of mycoplasmas detected by PCR was 57.14% in younger dogs (≤ 1 year) and 18.52% to 35.29% in older dogs, depending on year of the sampling. Genital swabs were positive in more cases (60%) in comparison with oral swabs (46.67%). M. canis was detected in 40% of positive cases, in the same percent of samples mixed not determined mycoplasma infections were confirmed. Mycoplasma species such as:  M. cynos, M. edwardii, M. maculosum, M. spumans were determined each in single cases and in one case mixed ureaplasma infection was confirmed. MOLEKULARNA DETEKCIJA IN SEROPREVALENCA MIKOPLAZEM PRI KLINIČNO ZDRAVIH DELOVNIH PSIHNamen raziskave je bil določiti seroprevalenco in prevalenco mikoplazem pri klinično zdravih delovnih psih. V raziskavo je bilo vključenih 34 delovnih psov različnih pasem in starosti, od tega 27 psov iz Slovenske vojske in 7 ovčarskih psov. Za dokazovanje specifičnih protiteles proti bakterijam Mycoplasma cynos, Mycoplasma canis in Mycoplasma molare smo uporabili metodo točkastega imunskega odtisa (ang. Dot Immuno Binding Assay- DIBA) in konvencionalni PCR, ki temelji na pomnoževanju odseka gena za ribosomalno RNK 16s ali intergenskega odseka genoma med genoma ribosomalnih RNK 16s in 23s. Specifična protitelesa proti vsaj eni od izbranih vrst mikoplazem so bila ugotovljena pri 94.1 % psov. Med njimi je 23.5 % vzorcev reagiralo pozitivno samo na M. canis, 20.6 % samo na M. canis in noben od vzorcev ni reagiral pozitivno samo na M. molare. Skupno je 47.0 % vzorcev reagiralo pozitivno na M. canis in M. canis hkrati, en pes (2.9 %) je imel specifična protitelesa proti vsem trem testiranim mikoplazmam. Z metodo PCR smo mikoplazme dokazali v vzorcih 57.1 % psov mlajših od enega leta, in pri 18.5 % do 35.3 % starejših od enega leta, odvisno od leta vzorčenja. Genitalni brisi so bili pozitivni v 60 % primerov v primerjavi z oralnimi kjer je bil delež 46.7 %. M. canis je bila ugotovljena v 40 % pozitivnih primerov, v enakem deležu so bile ugotovljene tudi mešane nedeterminirane mikoplazemske okužbe. Mikoplazme, kot so M. cynos, M. edwardii, M. maculosumin, M. spumans so bile ugotovljene posamično. V enem primeru je bila ugotovljena mešana okužba z ureaplazmami.Ključne besede: delovni psi; pasje mikoplazme; Mycoplasma canis; Mycoplasma cynos; DIBA; PCR