Table 1.
Materials used for HFF-1 cell culture and microsporidian spore isolation.
Fig 1.
Human foreskin fibroblast (HFF-1) cell culture monolayer.
HFF-1 cells 100% confluent observed under phase-contrast inverted microscope with a 40X magnification and the Canon EOS REBEL T3i digital camera.
Fig 2.
Encephalitozoon intestinalis ATCC 50506 spores in HFF-1 cell culture.
Culture visualized under phase-contrast inverted microscope with 10X objective and Canon EOS REBEL T3i digital camera. Each spore is about 1 µm wide to 3.5 µm in length.
Fig 3.
HFF-1 cells infected with Encephalitozoon hellem ATCC 50451 at different post- infection days. E. hellem vacuoles in HFF-1 cell culture (magenta arrow) at 7 dpi (A), 27 dpi (B), and 32 dpi (harvest stage) (C).
Fig 4.
Encephalitozoon cuniculi ATCC 50602 purified spores.
Single spores are marked by magenta arrows. Spore suspension was diluted to a 1:100 ratio and visualized/counted in hemocytometer under phase-contrast microscope with 10X objective and Canon EOS REBEL T3i digital camera.
Table 2.
Primers [22] used to amplify bacterial, mammal, and microsporidian DNA.
Fig 5.
Amplification of bacterial 16S, mammal and microsporidian rRNA in the E. intestinalis ATCC 50506 spore sample obtained from HFF-1 cell culture.
Ladder: 100 bp ladder (FroggaBio, ON, Canada). Neg: negative control, no sample. Well 1: E. intestinalis/E. hellem lsuF1-lsuR1 primers. Well 2: E. intestinalis/E. hellem lsuF2-lsuR2 primers. Well 3: E. intestinalis/E. hellem ssuF1-ssuR1 primers. Well 4: bacterial 357F-1100R primers. Well 5: mammal MF787- MR7535 primers. Bac + : Staphylococcus salivarius as positive control for bacteria primers.
Fig 6.
PCR of gDNA sample extracted from E. intestinalis ATCC 50506 spores purified from infected HFF-1 cells post DNase I treatment.
Ladder: 100 bp ladder. Neg: negative control with all used primers. Well 1: universal bacterial primers for 16S gene (357F-1100R). Well 2: universal mammal primers for rRNA gene (MF787-MR7535). Wells 3-5: E. intestinalis specific primers for rRNA (lsuF1-lsuR1, lsuF2-lsuR2, and ssuF1-ssuR1). Well 6: universal microsporidian primers for small rRNA subunit (ss530F- ss1047R).
Fig 7.
Estimations of gel band proportions with ImageJ.
The proportions estimated for bands in electrophoresis for PCR of human (H), bacterial (B) and microsporidian (M) DNA in the extracted gDNA sample.
Table 3.
Proportion of sequencing reads that mapped or did not map to the complete E. intestinalis ATCC 50506 genome.