Cryptosporidium spp. in caged exotic psittacines from Brazil: Evaluation of diagnostic methods and molecular characterization

Author(s)

Ferrari, Elis Domingos ^[1] ; Nakamura, Alex Akira ^[1] ; Nardi, Ana Rita Moraes ^[2] ; Santana, Bruna Nicoleti ^[1] ; Camargo, Vinicius da Silva ^[1] ; Nagata, Walter Bertequini ^[1] ; Saraiva Bresciani, Katia Denise ^[1] ; Meireles, Marcelo Vasconcelos ^[1]

Affiliation

^[1] Univ Estadual Paulista Unesp, Fac Med Vet, Clovis Pestana St 793, BR-16050680 Aracatuba, SP - Brazil

^[2] Fundacao Municipal Ensino Super, Estevao Diamant St 210, BR-12929590 Braganca Paulista, SP - Brazil

• Document type: Journal article
• Source: Experimental Parasitology; v. 184, p. 109-114, JAN 2018.
• Web of Science Citations: 3

Abstract

The aim of this study was to evaluate the prevalence of and diagnostic methods for Cryptosporidium spp. in caged adult exotic parrots from Southern and Southeastern Brazil. Oocysts were purified from fecal samples from 463 psittacines by centrifugal-flotation in Sheather's sugar solution. Cryptosporidium spp. were detected by malachite green negative staining and nested PCR targeting the 18S rRNA gene. Cryptosporidium species were identified by sequencing nested PCR amplicons. Samples were also tested by duplex real-time PCR targeting the 185 rRNA gene of Cryptosporidium galli and Cryptosporidium avian genotype III. The prevalence rates of Cryptosporidium spp. determined by microscopy and nested PCR were 3.0% (14/463) and 5.0% (23/463), respectively. The nested PCR/sequencing identified avian genotype III (1.7%; 8/463), Cryptosporidium parvum (0.9%; 4/463) and Cryptosporidium canis (0.2%; 1/463). Duplex real-time PCR was positive for gastric Cryptosporidium in 9.5% (44/463) of the samples. Among them, 1.9% (9/463) were positive for C. galli, 5.8% (27/463) were positive for avian genotype Ill and 1.7% (8/463) showed mixed infections with C galli and avian genotype Ill. With regards to the positive detection of Cryptosporidium spp., there was no statistically significant difference between nested PCR and microscopic analysis (p = .1237), and a fair agreement existed between them (Kappa = 0.242). A statistically significant difference (p < .0001) and fair agreement (Kappa = 0.317) were obtained between nested PCR/sequencing and duplex real-time PCR for the detection of gastric Cryptosporidium. We determined that nested PCR and duplex real-time PCR are the best options for the detection of Cryptosporidium spp. and gastric Cryptosporidium, respectively, and that avian genotype III is the most common Cryptosporidium genotype/species in psittacines. (C) 2017 Elsevier Inc. All rights reserved. (AU)

• FAPESP's process: 15/26334-8 - Cryptosporidium spp. in domestic birds and wild birds in the South and Southeast Brazil: occurrence, evaluation of methods of diagnosis and molecular characterization
• Grantee: Marcelo Vasconcelos Meireles
• Support Opportunities: Regular Research Grants