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 ZAVETISCA Na kliniko smo zaradi akutne krvave, penaste in smrdljive driske sprejeli tri mesece starega psa, ki so ga lastniki nedavno posvojili iz zavetisca. Klinicni pregled ter hematoloske in biokemicne preiskave krvi so bili brez posebnosti. Pri pregledu trebuha z ultrazvokom smo ugotovili velike mezenterialne bezgavke, tanko crevo pa je bilo polno ehogene vsebine. Z mikroskopskim pregledom nativnih preparatov smo v vzorcu blata opazili stevilne gibljive organizme, podobne trihomonasom, s specificnim krožnim gibanjem. Z mikroskopiranjem ter s flotacijo in metodo SAF smo diagnosticirali se jajceca nematodov Toxocara canis in oociste praživali Isospora sp. Trihomonasom podobne organizme smo uspesno izolirali in jih vzgojili v cisti kulturi. Mikroskopski pregled organizmov, obarvanih po Giemsi, je omogocil dolocitev stevila bickov in vzpostavitev suma na okužbo s Pentatrichomonas hominis (P. hominis). Diagnozo smo dokoncno potrdili s polimerazno verižno reakcijo (PCR) in dolocitvijo baznega zaporedja, ter z vrsticnim elektronskim mikroskopom. PCR in dolocitev baznega zaporedja sta pokazala 99 % skladnost nasega izolata z izolati P. hominis iz drugih gostiteljev/studij. Ta podatek kaže na zoonotski potencial P. hominis in na možnost peroralnega prenosa med živalmi in ljudmi. Clanek predstavlja tudi prvo porocilo o trihomonadni okužbi s P. hominis pri psih v Sloveniji. Kljucne besede: Pentatrichomonas hominis; pes; izolacija; gojenje; PCR; dolocanje zaporedja DNK; vrsticna elektronska mikroskopija