Encephalitozoon cuniculi (EC) is one of the most common and extensively studied microsporidian pathogens in rabbits, although infections have also been described in other mammalian species. Due to the growing body of research on its prevalence, clinical presentation, pathogenesis, and zoonotic potential, EC is considered an emerging pathogen in veterinary medicine. Our findings indicate that Slovenia has a high seroprevalence of EC, as 71.4% of rabbits are testing positive for EC-specific antibodies. In addition to the well-recognized forms of encephalitozoonosis affecting the central nervous system, urinary tract, and eyes, we identified and described a previously unreported atypical manifestation of EC infection in rabbits, namely gastrointestinal encephalitozoonosis, which was observed in nearly one-third of the studied animals. Furthermore, in rabbits with urinary and neurological forms of the disease, we statistically confirmed the occurrence of a markedly distended atonic urinary bladder. Using polymerase chain reaction (PCR), we found that 14.98% of the examined rabbits shed EC DNA into the environment through urine during clinical examination, hospitalization, or handling. Given the zoonotic potential of EC, these findings are important for veterinary personnel and rabbit owners. In addition, by analyzing non-standard samples (swabs from various mucosal surfaces and fur, as well as blood samples), we assessed contamination of rabbits with EC DNA and found it to be low (6.31%). This suggests that transmission to humans and other animals through contact with excretions or petting an infected rabbit is unlikely.
A low prevalence of EC in urine, blood, and feces (1/267; 0.34%) was also detected in other species of pet small mammals. All EC genotypes identified in rabbits and small mammals belonged to genotype I. Finally, we evaluated whether urinary biochemical analysis using a reagent rotor system could predict selected blood biochemical parameters; however, this approach proved to have limited clinical utility. Nevertheless, additional investigation of urine biochemistry confirmed that elevated urinary gamma-glutamyl transferase (GGT) may serve as an early indicator of kidney disease in rabbits.
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