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1
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Application of specific primers in the diagnosis of Encephalitozoon spp.

100%
Valencakova A., Balent P., Novotny F., Cislakova L.
Annals of Agricultural and Environmental Medicine
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2005
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tom 12
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nr 2
321-323
In our experiment, 3 species-specific primer pairs cultivated in cell lines were used: Encephalitozoon cuniculi-specific primer pairs (ECUNF and ECUNR), Encephalitozoon hellem-specific primer pairs (EHELF and EHELR), and Encephalitozoon intestinalis-specific primer pairs (SINTF and SINTR). The PCR products were estimated to be 550 bp in E. cuniculi, 547 bp in E. hellem and 545 bp in E. intestinalis respectively, which can prove the precision and reliability of this method in the species identification of the genus Encephalitozoon. All 3 primer pairs were species-specific and none of them amplified gene sequences from other Encephalitozoon spp.
2
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Prevalence of antibodies to Encephalitozoon cuniculi in horses in Israel

100%
Levkutova M., Hipikova V., Faitelzon S., Benath G., Paulik S., Levkut M.
Annals of Agricultural and Environmental Medicine
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2004
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tom 11
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nr 2
Infection with the intracellular microsporidium Encephalitozoon cuniculi can cause a serious disease - encephalitozoonosis in various animals and people. Several species of mammals, including the horse, were seem to be potential sources of encephalitozoonosis for animal as well as human hosts. The disease diagnosis is based on clinical signs, pathological findings, and the detection of E. cuniculi or circulating antibodies directed against the parasite. This study investigates the seroconversion to E. cuniculi in horses admitted to the Veterinary Teaching Hospital of the Hebrew University of Jerusalem and 3 different private horse-riding farms across Israel. Antibodies to E. cuniculi were determined using the IFA test in the sera from 102 horses. Of 72 asymptomatic horses, 60% were seropositive and 19% of the positive samples showed a titter of 1:512. Of 30 horses with various clinical signs, 80% were seropositive and 68% of the positive samples showed a titer of 1:512. High titers were associated with colic and neurological signs. This could prove to be interesting if the high percentages of prevalence of antibodies level in horses are an indication of health risk in humans.
3

Prevalence of anti-microsporidial antibodies in randomly examined dogs in Eastern Slovakia

100%
Herich R., Levkutova M., Kokincakova T., Reiterova K., Hipikova V., Levkut M.
Medycyna Weterynaryjna
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2008
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tom 64
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nr 05
Sixty-eight dogs were examined for the presence of Encephalitozoon spp. antibodies. Twenty-one dogs (30.9%) were healthy without any clinical signs of diseases. Forty-seven animals (69.1%) developed clinical symptoms of diseases such as chronic otitis externa, conjunctivitis, upper respiratory tract inflammation, status epilepticus, pyodermatitis, skin hypersensitivity, demodicosis, flea allergy. Different detection methods of encephalitozoonosis including IFAT (Indirect Immunofluorescence Antibody Assay), in vitro cultivation, SDS PAGE electrophoresis, Western blot and PCR were applied. There were 33 (48.5%) positive reacting sera to E. cuniculi II (mouse type) antigen using IFAT, including 9 positive samples obtained from clinically healthy dogs. Sixteen samples with the antibody titer equal to 1 : 256 were then tested by Western Blot. Most of the samples reacted with E. cuniculi II and III antigens. The presence of E. intestinalis antibodies was lower and just a few samples reacted with E. hellem antigen. The electrophoretic analysis of the encephalitozoon strains used as antigens confirmed that they differ primarily in the molecular size. The strain of type II (mouse) expressed a double strip at 54 and 58 kDa level. The strain of type III (dog) expressed a wide strip at 59 kDa. E. cuniculi types II and III are more related in protein structure in comparison to the other analyzed strains. When PMP1/PMP2 primers were used in PCR, the size of the amplified product was 268 bp for E. cuniculi and 270 bp for E. intestinalis. A species-specific primer pair for E. cuniculi ECUNF/ECUNR gave a 549 bp fragment and V1/SI-500 primers specific for E. intestinalis gave a 370 bp fragment.
4
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Occurrence of Encephalitozoon intestinalis in the Red ruffed lemur (Varecia rubra) and the Ring-tailed lemur (Lemur catta) housed in the Poznan Zoological Garden, Poland

80%
Slodkowicz-Kowalska A., Majewska A. C., Trzesowska E., Skrzypczak L.
Annals of Parasitology
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2012
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tom 58
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nr 1
Encephalitozoon intestinalis is one of the most common microsporidial species found in humans worldwide but it has rarely been identified in animals. The presence of this pathogen has been detected in a few species of domestic, captive and wild mammals as well as in three species of birds. The aim of the present study was to examine fecal samples obtained from mammals housed in the Poznan Zoological Garden, Poland, for the presence of potentially human-infectious microsporidia. A total of 339 fresh fecal samples collected from 75 species of mammals belonging to 27 families and 8 orders were examined for the presence of microsporidian spores. Microsporidian spores were identified in 3 out of 339 (0.9%) examined fecal samples. All samples identified as positive by chromotrope 2R and calcofluor white M2R were also positive by the FISH assay. Using multiplex FISH in all 3 fecal samples, only spores of E. intestinalis were identified in 2 out of 14 Ring-tailed lemurs (Lemur catta) and in one out of 17 Red ruffed lemurs (Varecia variegata rubra). To our knowledge this is the first diagnosis of E. intestinalis in Ring-tailed and Red ruffed lemurs. It should be mentioned that both lemur species are listed by the IUCN Red List of Threatened Species. Although the lemurs were asymptomatically infected, the possibility of widespread infection or death of these animals remains in the event of an elevated stress or a decrease in their immunological functions.
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