Fig. 1.
Phylogenetic relationships of 32 nucleotide sequences of Cryptosporidium species isolated from pet birds. The analysis was performed using the neighbour-joining method based on nucleotide sequences of the small-subunit ribosomal RNA gene. The evolutionary distances were computed using the Kimura two-parameter method and are the numbers of base substitutions per site. Two-letter and six-digit codes are GenBank accession numbers. Toxoplasma gondii was used as the outgroup
Fig. 2.
Melting curves obtained during qPCR-high-resolution melting analysis of the products of amplification of isolates of Cryptosporidium spp. from the faeces of pet birds. The paired curves for C. meleagridis and C. proventriculi indicate technical replicates or distinct isolates. RFU – relative fluorescence units
Fig. 3.
Derivative melting curve analysis of the products of amplification of isolates of Cryptosporidium spp from the faeces of pet birds. The paired curves for C. meleagridis and C. proventriculi indicate technical replicates or distinct isolates. –dF/dT – negative first derivative of fluorescence with respect to temperature
Table 2.
Results of the PCR test for the presence of two Cryptosporidium spp. in the studied bird group
| Species | n | C. proventriculi | C. meleagridis | ||
|---|---|---|---|---|---|
| n (+) | % | n (+) | % | ||
| African grey parrot (Psittacus erithacus) | 3 | 2 | 66.7 | 1 | 33.3 |
| Blue-and-yellow macaw (Ara ararauna) | 2 | 0 | 0.0 | 0 | 0.0 |
| Blue-fronted amazon (Amazona aestiva) | 2 | 1 | 50.0 | 0 | 0.0 |
| Blue-headed pionus (Pionus menstruus) | 1 | 0 | 0.0 | 0 | 0.0 |
| Budgerigar (Melopsittacus undulatus) | 4 | 0 | 0.0 | 0 | 0.0 |
| Canary (Serinus canaria) | 3 | 0 | 0.0 | 0 | 0.0 |
| Cockatiel (Nymphicus hollandicus) | 20 | 6 | 30 | 0 | 0.0 |
| Eastern rosella (Platycercus eximius) | 2 | 0 | 0.0 | 0 | 0.0 |
| Moluccan eclectus (Eclectus roratus) | 1 | 1 | 100 | 0 | 0.0 |
| Golden-collared macaw (Primolius auricollis) | 1 | 1 | 100 | 0 | 0.0 |
| Green-cheeked conure (Pyrrhura molinae) | 3 | 1 | 33.3 | 0 | 0.0 |
| Peach-faced lovebird (Agapornis roseicollis) | 4 | 1 | 25 | 0 | 0.0 |
| Pigeon (Columba livia) | 8 | 0 | 0.0 | 5 | 62.5 |
| Plum-headed parakeet (Psittacula cyanocephala) | 2 | 0 | 0.0 | 0 | 0.0 |
| Rainbow Lorikeet (Trichoglossus moluccanus) | 2 | 0 | 0.0 | 0 | 0.0 |
| Ring-necked parakeet (Psittacula krameri) | 1 | 0 | 0.0 | 0 | 0.0 |
| Senegal parrot (Poicephalus senegalus) | 1 | 0 | 0.0 | 0 | 0.0 |
| Sun conure (Aratinga solstitialis) | 1 | 0 | 0.0 | 0 | 0.0 |
| Timneh grey parrot (Psittacus erithacus timneh) | 1 | 1 | 100 | 0 | 0.0 |
| Yellow-headed amazon (Amazona oratrix) | 1 | 1 | 100 | 0 | 0.0 |
| Total | 63 | 15 | 23.8 | 6 | 9.52 |