TABLE 1.
Analysis of water for microsporidia by IFA, PCR, and sequencinga
| Sample type | Result of:
|
Speciesb | Accession no.c | Scored | % Similaritye | Pf | |
|---|---|---|---|---|---|---|---|
| IFA | PCR | ||||||
| Raw sewage | + | + | Encephalitozoon (Septata) intestinalis | L39113 | 391 | 97 | 1e-107 |
| Tertiary effluentg | + | + | Encephalitozoon (Septata) intestinalis | L39113 | 418 | 98 | 1e-115 |
| Tertiary effluentg | + | + | V. corneae | gb U11046 | 376 | 98 | 1e-90 |
| Tertiary effluentg | + | − | nd | nd | nd | nd | nd |
| Surface waterh | + | − | nd | nd | nd | nd | nd |
| Surface waterh | + | + | Encephalitozoon (Septata) intestinalis | U09929 | 289 | 96 | 9e-77 |
| Surface waterh | nd | + | Encephalitozoon (Septata) intestinalis | U39113 | 391 | 97 | 1e-98 |
| Surface wateri | nd | + | Enterocytozoon bieneusi | L16868 | 363 | 98 | 6e-99 |
| Controlj | + | + | Encephalitozoon (Septata) intestinalis | U39297 | 307 | 98 | 2e-82 |
| Controlj | nd | + | nd | nd | nd | nd | nd |
| Groundwater | − | + | Encephalitozoon (Septata) intestinalis | U39297 | 365 | 96 | 1e-85 |
| Groundwaterk | + | − | nd | nd | nd | nd | nd |
| Groundwaterk | + | − | nd | nd | nd | nd | nd |
| Groundwaterk | + | − | nd | nd | nd | nd | nd |
| Groundwaterk | + | − | nd | nd | nd | nd | nd |
| Groundwaterk | + | − | nd | nd | nd | nd | nd |
Results of PCR sequencing and species determination of microsporidia by using computer database homology searching. This method matches the unknown sequence with its closest relative in sequence databases. Similar sequences are ranked in order of highest homology and given statistical scores based on this homology. nd, not determined.
Species with the highest homology to the unknown sequence.
Accession number of the sequence with highest homology to the unknown sequence.
Ranking score given to the sequence with highest homology.
Percent base-to-base similarity between unknown and homologous sequences. It should be noted that in almost every instance the bases missed were due to a call of N by the sequencing program.
P, probability (on a scale of 0 to 1) that the sequence identified occurred by chance.
Activated sludge followed by pressure filtration through mixed-medium filters and extended chlorination.
Impacted with tertiary wastewater effluent.
No impact with discharged domestic wastewater.
Control, groundwater samples seeded with Encephalitozoon intestinalis spores.
Potentially impacted by surface wastewater infiltration.