Abstract
The morphological, biological, and molecular characteristics of Cryptosporidium avian genotype V are described, and the species name Cryptosporidium avium is proposed to reflect its specificity for birds under natural and experimental conditions. Oocysts of C. avium measured 5.30–6.90 μm (mean = 6.26 μm) × 4.30–5.50 μm (mean = 4.86 μm) with a length to width ratio of 1.29 (1.14–1.47). Oocysts of C. avium obtained from four naturally infected red-crowned parakeets (Cyanoramphus novaezealandiae) were infectious for 6-month-old budgerigars (Melopsittacus undulatus) and hens (Gallus gallus f. domestica). The prepatent periods in both susceptible bird species was 11 days postinfection (DPI). The infection intensity of C. avium in budgerigars and hens was low, with a maximum intensity of 5000 oocysts per gram of feces. Oocysts of C. avium were microscopically detected at only 12–16 DPI in hens and 12 DPI in budgerigars, while PCR analyses revealed the presence of specific DNA in fecal samples from 11 to 30 DPI (the conclusion of the experiment). Cryptosporidium avium was not infectious for 8-week-old SCID and BALB/c mice (Mus musculus). Naturally or experimentally infected birds showed no clinical signs of cryptosporidiosis, and no pathology was detected. Developmental stages of C. avium were detected in the ileum and cecum using scanning electron microscopy. Phylogenetic analyses based on small subunit rRNA, actin, and heat shock protein 70 gene sequences revealed that C. avium is genetically distinct from previously described Cryptosporidium species.
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References
Abe N, Makino I (2010) Multilocus genotypic analysis of Cryptosporidium isolates from cockatiels, Japan. Parasitol Res 106:1491–1497. doi:10.1007/s00436-010-1810-5
Akiyoshi DE, Dilo J, Pearson C, Chapman S, Tumwine J, Tzipori S (2003) Characterization of Cryptosporidium meleagridis of human origin passaged through different host species. Infect Immun 71:1828–1832. doi:10.1128/IAI.71.4.1828-1832.2003
Alves M, Xiao LH, Sulaiman I, Lal AA, Matos O, Antunes F (2003) Subgenotype analysis of Cryptosporidium isolates from humans, cattle, and zoo ruminants in Portugal. J Clin Microbiol 41:2744–2747. doi:10.1128/JCM.41.6.2744-2747.2003
Bermudez AJ, Ley DH, Levy MG, Ficken MD, Guy JS, Gerig TM (1988) Intestinal and bursal cryptosporidiosis in turkeys following inoculation with Cryptosporidium sp. isolated from commercial poults. Avian Dis 32:445–450. doi:10.2307/1590910
Cama VA, Bern C, Sulaiman IM, Gilman RH, Ticona E, Vivar A, Kawai V, Vargas D, Zhou L, Xiao LH (2003) Cryptosporidium species and genotypes in HIV-positive patients in Lima, Peru. J Eukaryot Microbiol 50:531–533. doi:10.1111/j.1550-7408.2003.tb00620.x
Current WL, Upton SJ, Haynes TB (1986) The life cycle of Cryptosporidium baileyi n. sp. (Apicomplexa, Cryptosporidiidae) infecting chickens. J Protozool 33:289–296. doi:10.1111/j.1550-7408.1986.tb05608.x
Curtiss JB, Leone AM, Wellehan JF Jr, Emerson JA, Howerth EW, Farina LL (2015) Renal and cloacal cryptosporidiosis (Cryptosporidium avian genotype V) in a Major Mitchell’s Cockatoo (Lophochroa Leadbeateri). J Zoo Wildl Med 46:934–937. doi:10.1638/2015-0025.1
Darabus G, Olariu R (2003) The homologous and interspecies transmission of Cryptosporidium parvum and Cryptosporidium meleagridis. Pol J Vet Sci 6:225–228. doi:10.1371/journal.pone.0054857
Elwin K, Robinson G, Hadfield SJ, Fairclough HV, Iturriza-Gomara M, Chalmers RM (2012) A comparison of two approaches to extracting Cryptosporidium DNA from human stools as measured by a real-time PCR assay. J Microbiol Methods 89:38–40. doi:10.1016/j.mimet.2012.02.006
Fayer R (2010) Taxonomy and species delimitation in Cryptosporidium. Exp Parasitol 124:90–97. doi:10.1016/j.exppara.2009.03.005
Fayer R, Santín M, Xiao L (2005) Cryptosporidium bovis n. sp. (Apicomplexa: Cryptosporidiidae) in cattle (Bos taurus). J Parasitol 91:624–629. doi:10.1645/GE-3435
Fayer R, Santín M, Trout JM (2008) Cryptosporidium ryanae n. sp. (Apicomplexa: Cryptosporidiidae) in cattle (Bos taurus). Vet Parasitol 156:191–198. doi:10.1016/j.vetpar.2008.05.024
Gharagozlou MJ, Dezfoulian O, Rahbari S, Bokaie S, Jahanzad I, Razavi AN (2006) Intestinal cryptosporidiosis in turkeys in Iran. J Vet Med A Physiol Pathol Clin Med 53:282–285. doi:10.1111/j.1439-0442.2006.00843.x
Graczyk TK, Fayer R, Trout JM, Lewis EJ, Farley CA, Sulaiman I, Lal AA (1998) Giardia sp. cysts and infectious Cryptosporidium parvum oocysts in the feces of migratory Canada geese (Branta canadensis). Appl Environ Microbiol 64:2736–2738
Guindon S, Gascuel O (2003) A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol 52:696–704. doi:10.1080/10635150390235520
Hornok S, Bitay Z, Szell Z, Varga I (1998) Assessment of maternal immunity to Cryptosporidium baileyi in chickens. Vet Parasitol 79:203–212. doi:10.1016/S0304-4017(98)00170-8
Huang K, Akiyoshi DE, Feng X, Tzipori S (2003) Development of patent infection in immunosuppressed C57Bl/6 mice with a single Cryptosporidium meleagridis oocyst. J Parasitol 89:620–622. doi:10.1645/0022-3395(2003)089[0620:dopiii]2.0.co;2
Jellison KL, Distel DL, Hemond HF, Schauer DB (2004) Phylogenetic analysis of the hypervariable region of the 18S rRNA gene of Cryptosporidium oocysts in feces of Canada geese (Branta canadensis): evidence for five novel genotypes. Appl Environ Microbiol 70:452–458. doi:10.1128/AEM.70.1.452-458.2004
Jellison KL, Lynch AE, Ziemann JM (2009) Source tracking identifies deer and geese as vectors of human-infectious Cryptosporidium genotypes in an urban/suburban watershed. Environ Sci Technol 43:4267–4272. doi:10.1021/es900081m
Jiang J, Alderisio KA, Xiao L (2005) Distribution of Cryptosporidium genotypes in storm event water samples from three watersheds in New York. Appl Environ Microbiol 71:4446–4454. doi:10.1128/AEM.71.8.4446-4454.2005
Kilani RT, Sekla L (1987) Purification of Cryptosporidium oocysts and sporozoites by cesium chloride and Percoll gradients. Am J Trop Med Hyg 36:505–508
Kváč M, Vítovec J (2003) Prevalence and pathogenicity of Cryptosporidium andersoni in one herd of beef cattle. J Veterinary Med Ser B 50:451–457. doi:10.1046/j.0931-1793.2003.00701.x
Kváč M, Ondráčková Z, Květoňová D, Sak B, Vítovec J (2007) Infectivity and pathogenicity of Cryptosporidium andersoni to a novel host, southern multimammate mouse (Mastomys coucha). Vet Parasitol 143:229–233. doi:10.1016/j.vetpar.2006.08.031
Kváč M, Hofmannová L, Hlásková L, Květoňová D, Vítovec J, McEvoy J, Sak B (2014a) Cryptosporidium erinacei n. sp. (Apicomplexa: Cryptosporidiidae) in hedgehogs. Vet Parasitol 201:9–17. doi:10.1016/j.vetpar.2014.01.014
Kváč M, McEvoy J, Stenger B, Clark M (2014b) Cryptosporidiosis in other vertebrates. In: Cacciò SM, Widmer G (eds) Cryptosporidium: parasite and disease, 1st edn. Springer, Wien, pp 237–326
Lindsay DS, Blagburn BL (1990) Cryptosporidiosis in birds. In: Dubey JP, Speer CA, Fayer R (eds) Cryptosporidiosis in man and animals, 1st edn. CRC Press, Boca Raton, FL, pp 133–148
Lindsay DS, Blagburn BL, Sundermann CA, Giambrone JJ (1988) Effect of broiler chicken age on susceptibility to experimentally induced Cryptosporidium baileyi infection. Am J Vet Res 49:1412–1414
Liu X, He T, Zhong Z, Zhang H, Wang R, Dong H, Wang C, Li D, Deng J, Peng G, Zhang L (2013) A new genotype of Cryptosporidium from giant panda (Ailuropoda melanoleuca) in China. Parasitol Int 62:454–458. doi:10.1016/j.parint.2013.06.004
Makino I, Abe N, Reavill DR (2010) Cryptosporidium avian genotype III as a possible causative agent of chronic vomiting in peach-faced lovebirds (Agapornis roseicollis). Avian Dis 54:1102–1107. doi:10.1637/9227-123009-Case.1
McLauchlin J, Amar C, Pedraza-Diaz S, Nichols GL (2000) Molecular epidemiological analysis of Cryptosporidium spp. in the United Kingdom: results of genotyping Cryptosporidium spp. in 1705 fecal samples from humans and 105 fecal samples from livestock animals. J Clin Microbiol 38:3984–3990
Meireles MV, Soares RM, dos Santos MM, Gennari SM (2006) Biological studies and molecular characterization of a Cryptosporidium isolate from ostriches (Struthio camelus). J Parasitol 92:623–626. doi:10.1645/0022-3395(2006)92[623:bsamco]2.0.co;2
Miláček P, Vítovec J (1985) Differential staining of cryptosporidia by aniline-carbol-methyl violet and tartrazine in smears from feces and scrapings of intestinal mucosa. Folia Parasitol 32:50
Nakamura AA, Meireles MV (2015) Cryptosporidium infections in birds—a review. Rev Bras Parasitol Vet 24:253–267. doi:10.1590/S1984-29612015063
Nakamura AA, Homem CG, da Silva AM, Meireles MV (2014) Diagnosis of gastric cryptosporidiosis in birds using a duplex real-time PCR assay. Vet Parasitol 205:7–13. doi:10.1016/j.vetpar.2014.07.033
Ng J, Pavlásek I, Ryan U (2006) Identification of novel Cryptosporidium genotypes from avian hosts. Appl Environ Microbiol 72:7548–7553. doi:10.1128/AEM.01352-06
O’Donoghue PJ (1995) Cryptosporidium and cryptosporidiosis in man and animals. Int J Parasitol 25:139–195. doi:10.1016/0020-7519(94)E0059-V
Pavlásek I (2001) Findings of cryptosporidia in the stomach of hens and of exotic and wild birds. Veterinářství (Czech) 51:103–108
Qi M, Wang R, Ning C, Li X, Zhang L, Jian F, Sun Y, Xiao L (2011) Cryptosporidium spp. in pet birds: genetic diversity and potential public health significance. Exp Parasitol 128:336–340. doi:10.1016/j.exppara.2011.04.003
Rhee JK, Seu YS, Park BK (1991) Isolation and identification of Cryptosporidium from various animals in Korea. III. Identification of Cryptosporidium baileyi from Korean chicken. Korean J Parasitol 29:315–324. doi:10.3347/kjp.1991.29.4.315
Ryan U (2010) Cryptosporidium in birds, fish and amphibians. Exp Parasitol 124:113–120. doi:10.1016/j.exppara.2009.02.002
Ryan U, Xiao L, Read C, Zhou L, Lal AA, Pavlásek I (2003a) Identification of novel Cryptosporidium genotypes from the Czech Republic. Appl Environ Microbiol 69:4302–4307. doi:10.1128/AEM.69.7.4302-4307.2003
Ryan UM, Xiao L, Read C, Sulaiman IM, Monis P, Lal AA, Fayer R, Pavlásek I (2003b) A redescription of Cryptosporidium galli Pavlásek, 1999 (Apicomplexa : Cryptosporidiidae) from birds. J Parasitol 89:809–813. doi:10.1645/GE-74RI
Sauch JF, Flanigan D, Galvin ML, Berman D, Jakubowski W (1991) Propidium iodide as an indicator of Giardia cyst viability. Appl Environ Microbiol 57:3243–3247
Slavin D (1955) Cryptosporidium meleagridis (sp. nov.). J Comp Pathol 65:262–266
Sreter T, Varga I, Bekesi L (1995) Age-dependent resistance to Cryptosporidium baileyi infection in chickens. J Parasitol 81:827–829. doi:10.2307/3283992
Sulaiman IM, Morgan UM, Thompson RC, Lal AA, Xiao L (2000) Phylogenetic relationships of Cryptosporidium parasites based on the 70-kilodalton heat shock protein (HSP70) gene. Appl Environ Microbiol 66:2385–2391. doi:10.1128/AEM.66.6.2385-2391.2000
Sulaiman IM, Lal AA, Xiao LH (2002) Molecular phylogeny and evolutionary relationships of Cryptosporidium parasites at the actin locus. J Parasitol 88:388–394. doi:10.1645/0022-3395(2002)088[0388:MPAERO]2.0.CO;2
Tamura K (1992) Estimation of the number of nucleotide substitutions when there are strong transition-transversion and G + C-content biases. Mol Biol Evol 9:678–687
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739. doi:10.1093/molbev/msr121
Tavaré S (1986) Some probabilistic and statistical problems in the analysis of DNA sequences. In: Miura RM (ed) Some mathematical questions in biology: DNA sequence analysis (lectures on mathematics in the life sciences). American Mathematical Society, New York, pp 57–86
Tůmová E, Skřivan M, Marounek M, Pavlásek I, Ledvinka Z (2002) Performance and oocyst shedding in broiler chickens orally infected with Cryptosporidium baileyi and Cryptosporidium meleagridis. Avian Dis 46:203–207. doi:10.1637/0005-2086(2002)046[0203:PAOSIB]2.0.CO;2
Valigurová A, Jirku M, Koudela B, Gelnar M, Modrý D, Šlapeta J (2008) Cryptosporidia: epicellular parasites embraced by the host cell membrane. Int J Parasitol 38:913–922. doi:10.1016/j.ijpara.2007.11.003
Wang LM, Xue X, Li JQ, Zhou QJ, Yu YC, Du AF (2014) Cryptosporidiosis in broiler chickens in Zhejiang Province, China: molecular characterization of oocysts detected in fecal samples. Parasite. doi:10.1051/Parasite/2014035, 21
Woodmansee DB, Pavlásek I, Pohlenz JF, Moon HW (1988) Subclinical cryptosporidiosis of turkeys in Iowa. J Parasitol 74:898–900. doi:10.2307/3282279
Xiao L, Ryan UM (2004) Cryptosporidiosis: an update in molecular epidemiology. Curr Opin Infect Dis 17:483–490. doi:10.1097/00001432-200410000-00014
Xiao L, Morgan UM, Limor J, Escalante A, Arrowood M, Shulaw W, Thompson RC, Fayer R, Lal AA (1999) Genetic diversity within Cryptosporidium parvum and related Cryptosporidium species. Appl Environ Microbiol 65:3386–3391
Xiao L, Fayer R, Ryan U, Upton SJ (2004) Cryptosporidium taxonomy: recent advances and implications for public health. Clin Microbiol Rev 17:72–97. doi:10.1128/CMR.17.1.72-97.2004
Zhang XX, Zhang NZ, Zhao GH, Zhao Q, Zhu XQ (2015) Prevalence and genotyping of Cryptosporidium infection in pet parrots in North China. Biomed Res Int. doi:10.1155/2015/549798
Zhou L, Kassa H, Tischler ML, Xiao L (2004) Host-adapted Cryptosporidium spp. in Canada geese (Branta canadensis). Appl Environ Microbiol 70:4211–4215. doi:10.1128/aem.70.7.4211-4215.2004
Acknowledgments
This study was funded by the Czech Science Foundation (15-01090S) and by National Institutes of Health project 2P20 RR015566. Sarah Menchaca was supported in part by a grant of the University of Arizona from the National Institutes of Health (T37 MD001427).
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Holubová, N., Sak, B., Horčičková, M. et al. Cryptosporidium avium n. sp. (Apicomplexa: Cryptosporidiidae) in birds. Parasitol Res 115, 2243–2251 (2016). https://doi.org/10.1007/s00436-016-4967-8
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DOI: https://doi.org/10.1007/s00436-016-4967-8