Pentatrichomonas hominis COINFECTION IN A PUPPY FROM A SLOVENIAN ANIMAL SHELTER

A 3-month-old dog originating from a Slovenian animal shelter presented with acute bloody, soft, foamy and malodorous diarrhoea. The clinical examination, haematology and serum biochemistry were unremarkable. Ultrasonography of the abdomen showed prominent mesenteric lymph nodes and the presence of echogenic content within the small intestine. Light microscopy of a native smear and a wet mount darkfield microscopy examination of the faecal material showed motile trichomonad-like organisms with a particular circular motion. The flotation and SAF (Sodium acetate - acetic acid - formalin solution) method using light microscopy revealed eggs of nematode Toxocara canis and protozoan oocysts of Isospora spp. Trichomonad-like organisms were successfully isolated and cultivated in axenic culture. Light microscopy of Giemsa-stained trichomonads showed the presence of five flagella, and Pentatrichomonas hominis (P. hominis) was presumptively diagnosed. The diagnosis was confirmed by the Polymerase Chain Reaction (PCR) followed by DNA sequencing and the Scanning Electron Microscopy (SEM) of cultured trichomonad isolates. The PCR and sequencing results confirmed a 99% homology of the P. hominis isolates with isolates from other studies, originating both from humans and animals, which suggests that P. hominis could have zoonotic potential and have been transmitted from animals to people via the per-oral route. This is also the first report on P. hominis involvement in clinical diseases in dogs in Slovenia.Key words: Pentatrichomonas hominis; dog; isolation; cultivation; PCR; DNA sequencing; scanning electron microscopy SOOKUŽBA S PROTOZOJEM Pentatrichomonas hominis PRI PSU IZ SLOVENSKEGA ZAVETIŠČANa kliniko smo zaradi akutne krvave, penaste in smrdljive driske sprejeli tri mesece starega psa, ki so ga lastniki nedavno posvojili iz zavetišča. Klinični pregled ter hematološke in biokemične preiskave krvi so bili brez posebnosti. Pri pregledu trebuha z ultrazvokom smo ugotovili velike mezenterialne bezgavke, tanko črevo pa je bilo polno ehogene vsebine. Z mikroskopskim pregledom nativnih preparatov smo v vzorcu blata opazili številne gibljive organizme, podobne trihomonasom, s specifičnim krožnim gibanjem. Z mikroskopiranjem ter s flotacijo in metodo SAF smo diagnosticirali še jajčeca nematodov Toxocara canis in oociste praživali Isospora sp. Trihomonasom podobne organizme smo uspešno izolirali in jih vzgojili v čisti kulturi. Mikroskopski pregled organizmov, obarvanih po Giemsi, je omogočil določitev števila bičkov in vzpostavitev suma na okužbo s Pentatrichomonas hominis (P. hominis). Diagnozo smo dokončno potrdili s polimerazno verižno reakcijo (PCR) in določitvijo baznega zaporedja, ter z vrstičnim elektronskim mikroskopom. PCR in določitev baznega zaporedja sta pokazala 99 % skladnost našega izolata z izolati P. hominis iz drugih gostiteljev/študij. Ta podatek kaže na zoonotski potencial P. hominis in na možnost peroralnega prenosa med živalmi in ljudmi. Članek predstavlja tudi prvo poročilo o trihomonadni okužbi s P. hominis pri psih v Sloveniji.Ključne besede: Pentatrichomonas hominis; pes; izolacija; gojenje; PCR; določanje zaporedja DNK; vrstična elektronska mikroskopija