Abstract
This study used several diagnostic methods to examine the occurrence of and molecularly characterize Cryptosporidium spp. in captive canaries (Serinus canaria) in southern and southeastern Brazil. A total of 498 fecal samples were purified by centrifugal-flotation using Sheather's solution. Cryptosporidium spp. diagnosis was performed using three diagnostic methods: malachite green negative staining, nested PCR targeting the 18S rRNA gene, followed by sequencing the amplified fragments, and duplex real-time PCR targeting the 18S rRNA specific to detect Cryptosporidium galli and Cryptosporidium avian genotype III. The overall positivity for Cryptosporidium spp. (total samples positive in at least one protocol) from the microscopic analysis, nested PCR and duplex real-time PCR protocol results was 13.3% (66/498). The positivity rates were 2.0% (10/498) and 4.6% (23/498) for Cryptosporidium spp. by microscopy and nested PCR, respectively. Sequencing of 20 samples amplified by nested PCR identified C. galli (3.0%; 15/498), Cryptosporidium avian genotype I (0.8%; 4/498) and Cryptosporidium avium (0.2%; 1/498). Duplex real-time PCR revealed a positivity of 7.8% (39/498) for C. galli and 2.4% (12/498) for avian genotype III. Malachite green negative staining differed significantly from nested PCR in detecting Cryptosporidium spp. Duplex real-time PCR was more sensitive than nested PCR/sequencing for detecting gastric Cryptosporidium in canaries.
Highlights
Cryptosporidiosis is a major protozoan infection in birds, causing respiratory and gastrointestinal diseases in domestic and wild species (NAKAMURA & MEIRELES, 2015)
Sequencing of 20 samples amplified by nested PCR identified C. galli (3.0%; 15/498), Cryptosporidium avian genotype I (0.8%; 4/498) and Cryptosporidium avium (0.2%; 1/498)
Studies conducted in several countries, with several species of domestic and wild birds, reported that Cryptosporidium spp. prevalence ranged from 0.8% to 44.4%, and C. andersoni, C. avium, C. baileyi, C. galli, C. muris, C. meleagridis, and C. parvum and several avian genotypes, including the avian genotypes I, II, III and VI were identified (NAKAMURA & MEIRELES, 2015, CHELLADURAI et al, 2016)
Summary
Cryptosporidiosis is a major protozoan infection in birds, causing respiratory and gastrointestinal diseases in domestic and wild species (NAKAMURA & MEIRELES, 2015). In addition to the avian Cryptosporidium species, many genotypes infect birds, mainly the avian genotypes I (NG et al, 2006), II (SANTOS et al, 2005; MEIRELES et al, 2006; NG et al, 2006), and III (NG et al, 2006). Studies conducted in several countries, with several species of domestic and wild birds, reported that Cryptosporidium spp. prevalence ranged from 0.8% to 44.4%, and C. andersoni, C. avium, C. baileyi, C. galli, C. muris, C. meleagridis, and C. parvum and several avian genotypes, including the avian genotypes I, II, III and VI were identified (NAKAMURA & MEIRELES, 2015, CHELLADURAI et al, 2016). C. galli or Cryptosporidium avian genotype III infections can result in diarrhea, anorexia, weight loss, and chronic vomiting (ANTUNES et al, 2008; MAKINO et al, 2010, SILVA et al, 2010; RAVICH et al, 2014)
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