Abstract
Giardia duodenalis and Cryptosporidium spp. are common intestinal pathogens of humans and animals. Dogs may be infected by zoonotic isolates of G. duodenalis and Cryptosporidium spp. that, consequently, have high interest under public health perspective. This study estimated the occurrence of these protozoa in canine faeces polluting public areas of Padua municipality (Northern Italy), towards a potential evaluation of health risks for dogs and humans. A total of 705 canine stools was collected in green (n = 270) and urban (n = 435) areas and processed by duplex real-time PCR and real-time PCR SYBR® Green I for the detection of both protozoa. Positive samples were submitted to specific nested PCRs (i.e. β-giardin/SSU-rRNA genes for Giardia; SSU-rRNA gene for Cryptosporidium) to obtain detailed information on the isolates retrieved.Giardia and Cryptosporidium prevalence were 28.9% and 1.7%, respectively. Twenty-one Giardia-positive samples were successfully identified as dog-specific assemblages C and D, and 1 as the human-specific assemblage B. One isolate was identified as Cryptosporidium canis, while the other 11 were confirmed to belong to the Cryptosporidium parvum species complex. Contrariwise to the Cryptosporidium low prevalence, the wide distribution of Giardia suggests a high risk of infection for dogs attending public areas. Although data indicate a limited risk for human health, it is necessary to improve general education to reduce canine faecal pollution towards a widespread awareness of health risks.
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References
Adamska M, Leońska-Duniec A, Maciejewska A, Sawczuk M, Skotarczak B (2010) Comparison of efficiency of various DNA extraction methods from cysts of Giardia intestinalis measured by PCR and TaqMan real time PCR. Parasite 17(4):299–305. https://doi.org/10.1051/parasite/2010174299
Ballweber LR, Xiao L, Bowman DD et al (2010) Giardiasis in dogs and cats: update on epidemiology and public health significance. Trends Parasitol 26:180–189. https://doi.org/10.1016/j.pt.2010.02.005
Beraldo P, Candusso S, Mingotto F, Arzese A (2014) Canine faecal contamination in Udine and evaluation of health risk. In: Proceedings XXVIII National Congress Società Italiana di Parassitologia (SoIPa). p 258
Berrilli F, Di Cave D, De Liberato C et al (2004) Genotype characterisation of Giardia duodenalis isolates from domestic and farm animals by SSU-rRNA gene sequencing. Vet Parasitol 122:193–199. https://doi.org/10.1016/j.vetpar.2004.04.008
Bouzid M, Halai K, Jeffreys D, Hunter PR (2015) The prevalence of Giardia infection in dogs and cats, a systematic review and meta-analysis of prevalence studies from stool samples. Vet Parasitol 207:181–202. https://doi.org/10.1016/j.vetpar.2014.12.011
Bowman DD, Lucio-Forster A (2010) Cryptosporidiosis and giardiasis in dogs and cats: veterinary and public health importance. Exp Parasitol 124:121–127. https://doi.org/10.1016/j.exppara.2009.01.003
Cacciò SM, De Giacomo M, Pozio E (2002) Sequence analysis of the β-giardin gene and development of a polymerase chain reaction-restriction fragment length polymorphism assay to genotype Giardia duodenalis cysts from human faecal samples. Int J Parasitol 32:1023–1030. https://doi.org/10.1016/S0020-7519(02)00068-1
Cama VA, Bern C, Sulaiman IM et al (2003) Cryptosporidium species and genotypes in HIV-positive patients in Lima, Peru. J Eukaryot Microbiol 50(Suppl):531–533
Claerebout E, Casaert S, Dalemans AC et al (2009) Giardia and other intestinal parasites in different dog populations in Northern Belgium. Vet Parasitol 161:41–46. https://doi.org/10.1016/j.vetpar.2008.11.024
Coelho W, do Amarante AF, Apolinário J et al (2011) Occurence of Ancylostoma in dogs, cats, and public palces from Andradina city, Sao Paulo State, Brazil. Rev Inst Med Trop Sao Paulo 53:181–184. https://doi.org/10.1590/S0036-46652011000400001
Coronato B, Bastos O, Duarte R et al (2012) Parasites in stool samples in the environment of Ilha da Marambaia, Rio de Janeiro, Brazil: an approach in public health. Rev Inst Med Trop Sao Paulo 54:65–68. https://doi.org/10.1590/S0036-46652012000200002
Covacin C, Aucoin DP, Elliot A, Thompson RCA (2011) Genotypic characterisation of Giardia from domestic dogs in the USA. Vet Parasitol 177:28–32. https://doi.org/10.1016/j.vetpar.2010.11.029
Dado D, Izquierdo F, Vera O et al (2012a) Detection of zoonotic intestinal parasites in public parks of Spain. Potential epidemiological role of microsporidia. Zoonoses Public Health 59:23–28. https://doi.org/10.1111/j.1863-2378.2011.01411.x
Dado D, Montoya A, Blanco MA et al (2012b) Prevalence and genotypes of Giardia duodenalis from dogs in Spain: possible zoonotic transmission and public health importance. Parasitol Res 111:2419–2422. https://doi.org/10.1007/s00436-012-3100-x
Dubná S, Langrová I, Jankovská I et al (2007) Contamination of soil with Toxocara eggs in urban (Prague) and rural areas in the Czech Republic. Vet Parasitol 144:81–86. https://doi.org/10.1016/j.vetpar.2006.09.023
Epe C, Rehkter G, Schnieder T et al (2010) Giardia in symptomatic dogs and cats in Europe-results of a European study. Vet Parasitol 173:32–38. https://doi.org/10.1016/j.vetpar.2010.06.015
Feng Y, Xiao L (2011) Zoonotic potential and molecular epidemiology of Giardia species and giardiasis. Clin Microbiol Rev 24:110–140. https://doi.org/10.1128/CMR.00033-10
Genchi M, Ferroglio E, Traldi G et al (2007) Fecalizzazione ambientale e rischio parassitario nelle città di Milano e Torino. La Prof Vet:15–17
Giacometti A, Cirioni O, Fortuna M et al (2000) Environmental and serological evidence for the presence of toxocariasis in the urban area of Ancona, Italy. Eur J Epidemiol 16:1023–1026
Giangaspero A, Iorio R, Paoletti B, Traversa D, Capelli G (2006) Molecular evidence for in Central Italy. Parasitol Res 99:297–299. https://doi.org/10.1007/s00436-006-0169-0
Giangaspero A, Berrilli F, Brandonisio O (2007) Giardia and Cryptosporidium and public health: the epidemiological scenario from the Italian perspective. Parasitol Res 101:1169–1182. https://doi.org/10.1007/s00436-007-0598-4
Guy RA, Payment P, Krull UJ, Horgen PA (2003) Real-time PCR for quantification of Giardia and Cryptosporidium in environmental water samples and sewage. Appl Environ Microbiol 69:5178–5185. https://doi.org/10.1128/AEM.69.9.5178
Habluetzel A, Traldi G, Ruggieri S et al (2003) An estimation of Toxocara canis prevalence in dogs, environmental egg contamination and risk of human infection in the Marche region of Italy. Vet Parasitol 113:243–252. https://doi.org/10.1016/S0304-4017(03)00082-7
Heyworth MF (2016) Giardia duodenalis genetic assemblages and hosts. Parasite 23:13. https://doi.org/10.1051/parasite/2016013
Inpankaew T, Schär F, Odermatt P et al (2014) Low risk for transmission of zoonotic Giardia duodenalis from dogs to humans in rural Cambodia. Parasit Vectors 7:412. https://doi.org/10.1186/1756-3305-7-412
Itoh N, Kanai K, Hori Y et al (2009) Prevalence of Giardia intestinalis and other zoonotic intestinal parasites in private household dogs of the Hachinohe area in Aomori prefecture, Japan in 1997, 2002 and 2007. J Vet Sci 10:305–308. https://doi.org/10.4142/jvs.2009.10.4.305
Joffe D, Van Niekerk D, Gagné F et al (2011) The prevalence of intestinal parasites in dogs and cats in Calgary, Alberta. Can Vet J 52:1323–1328
Lalle M, Pozio E, Capelli G et al (2005) Genetic heterogeneity at the beta-giardin locus among human and animal isolates of Giardia duodenalis and identification of potentially zoonotic subgenotypes. Int J Parasitol 35:207–213. https://doi.org/10.1016/j.ijpara.2004.10.022
Leonhard S, Pfister K, Beelitz P et al (2007) The molecular characterisation of Giardia from dogs in Southern Germany. Vet Parasitol 150:33–38. https://doi.org/10.1016/j.vetpar.2007.08.034
Lia R, La Montanara C, Leone N, et al (2002) Canine helmintic fauna and environmental faecalization in the town of Bari (Apulia Region, Southern Italy). In: Proceedings XXII Nazional Congress Società Italiana di Parassitologia (SoIPa). p 92
Lucio-Forster A, Griffiths JK, Cama VA et al (2010) Minimal zoonotic risk of cryptosporidiosis from pet dogs and cats. Trends Parasitol 26:174–179. https://doi.org/10.1016/j.pt.2010.01.004
Macpherson CNL (2005) Human behaviour and the epidemiology of parasitic zoonoses. Int J Parasitol 35:1319–1331. https://doi.org/10.1016/j.ijpara.2005.06.004
MAFF (1986) Manual of veterinary parasitological laboratory techniques, 3rd edn. HMSO, London
Marangi M, Berilli F, Otranto D et al (2010) Genotyping of Giardia duodenalis among children and dogs in a closed socially deprived community from Italy. Zoonoses Public Health 57:e54–e58. https://doi.org/10.1111/j.1863-2378.2009.01304.x
Marques J, Guimarães C, Vilas Boas A et al (2012) Contamination of public parks and squares from Guarulhos (São Paulo State, Brazil) by Toxocara spp. and Ancylostoma spp. Rev Inst Med Trop Sao Paulo 54:267–271. https://doi.org/10.1590/S0036-46652012000500006
Monis PT, Caccio SM, Thompson RCA (2009) Variation in Giardia: towards a taxonomic revision of the genus. Trends Parasitol 25:93–100. https://doi.org/10.1016/j.pt.2008.11.006
Olson ME, Goh J, Phillips M, Guselle N, McAllister TA (1999) Giardia cyst and Cryptosporidium oocyst survival in water, soil, and cattle feces. J Environ Qual 28:1991–1996
Osman M, Bories J, El Safadi D et al (2015) Prevalence and genetic diversity of the intestinal parasites Blastocystis sp. and Cryptosporidium spp. in household dogs in France and evaluation of zoonotic transmission risk. Vet Parasitol 30; 214(1–2):167–170. https://doi.org/10.1016/j.vetpar.2015.09.015
Padua Municipality (2016) Popolazione. http://www.padovanet.it/informazione/padova-cifre. Accessed 6 July 2017
Palmer CS, Thompson RCA, Traub RJ et al (2008) National study of the gastrointestinal parasites of dogs and cats in Australia. Vet Parasitol 151:181–190. https://doi.org/10.1016/j.vetpar.2007.10.015
Paoletti B, Otranto D, Weigl S et al (2011) Prevalence and genetic characterization of Giardia and Cryptosporidium in cats from Italy. Res Vet Sci 91:397–399. https://doi.org/10.1016/j.rvsc.2010.09.011
Papini R, Marangi M, Mancianti F, Giangaspero A (2009) Occurrence and cyst burden of Giardia duodenalis in dog faecal deposits from urban green areas: implications for environmental contamination and related risks. Prev Vet Med 92:158–162. https://doi.org/10.1016/j.prevetmed.2009.07.003
Pedraza-Díaz S, Amar C, Iversen AM et al (2001) Unusual cryptosporidium species recovered from human faeces: first description of Cryptosporidium felis and Cryptosporidium “dog type” from patients in England. J Med Microbiol 50:293–296
Riggio F, Mannella R, Ariti G, Perrucci S (2013) Intestinal and lung parasites in owned dogs and cats from central Italy. Vet Parasitol 193:78–84. https://doi.org/10.1016/j.vetpar.2012.11.026
Rinaldi L, Biggeri A, Carbone S et al (2006) Canine faecal contamination and parasitic risk in the city of Naples (southern Italy). BMC Vet Res 2:29. https://doi.org/10.1186/1746-6148-2-29
Rinaldi L, Maurelli MP, Musella V et al (2008) Giardia and Cryptosporidium in canine faecal samples contaminating an urban area. Res Vet Sci 84:413–415. https://doi.org/10.1016/j.rvsc.2007.05.006
Risitano AL, Brianti E, Gaglio G, et al (2007) Environmental contamination by canine feces in the city of Messina: parasitological aspects and zoonotic hazards. In: Proceedings LXI National Congress Società Italiana delle Scienze Veterinarie (SISVet). pp 135–136
Robertson ID, Thompson RCA (2002) Enteric parasitic zoonoses of domesticated dogs and cats. Microbes Infect 4:867–873
Rossle NF, Latif B (2013) Cryptosporidiosis as threatening health problem: a review. Asian Pac J Trop Biomed 3(11):916–924. https://doi.org/10.1016/S2221-1691(13)60179-3
Rubel D, Wisnivesky C (2010) Contaminacion fecal canina en plazas y veredas de Buenos Aires, 1991-2006. Med (Buenos Aires) 70:355–363
Ryan U, Cacciò SM (2013) Zoonotic potential of Giardia. Int J Parasitol 43:943–956. https://doi.org/10.1016/j.ijpara.2013.06.001
Savilla TM, Joy JE, May JD, Somerville CC (2011) Prevalence of dog intestinal nematode parasites in south central West Virginia, USA. Vet Parasitol 178:115–120. https://doi.org/10.1016/j.vetpar.2010.12.034
Scala A, Garippa G, Pintus D (2009) Environmental contamination by canine feces in the city of Alghero (SS): parasitological aspects and zoonotic hazards. In: Proceedings LXIII National Congress Società Italiana delle Scienze Veterinarie (SISVet). pp 180–182
Scorza V, Tangtrongsup S (2010) Update on the diagnosis and management of Cryptosporidium spp. infections in dogs and cats. Top Companion Anim Med 25:163–169. https://doi.org/10.1053/j.tcam.2010.07.007
Simonato G, Frangipane di Regalbono A, Cassini R et al (2015) Copromicroscopic and molecular investigations on intestinal parasites in kenneled dogs. Parasitol Res 114:1963–1970. https://doi.org/10.1007/s00436-015-4385-3
Šlapeta J (2013) Cryptosporidiosis and Cryptosporidium species in animals and humans: a thirty colour rainbow? Int J Parasitol 43:957–970. https://doi.org/10.1016/j.ijpara.2013.07.005
Soliman RH, Fuentes I, Rubio JM (2011) Identification of a novel assemblage B subgenotype and a zoonotic assemblage C in human isolates of Giardia intestinalis in Egypt. Parasitol Int 60:507–511. https://doi.org/10.1016/j.parint.2011.09.006
Sotiriadou I, Pantchev N, Gassmann D, Karanis P (2013) Molecular identification of Giardia and Cryptosporidium from dogs and cats. Parasite 20:8. https://doi.org/10.1051/parasite/2013008
Stull JW, Peregrine AS, Sargeant JM, Weese JS (2012) Household knowledge, attitudes and practices related to pet contact and associated zoonoses in Ontario, Canada. BMC Public Health 12:553. https://doi.org/10.1186/1471-2458-12-553
Tamponi C, Varcasia A, Del Giudice F, et al (2014) Environmental contamination by pet faeces and parasitic risk in the city of Sassari (Sardinia, Italy). In: Proceedings XXVIII National Congress Società Italiana di Parassitologia (SoIPa). p 257
Tangtrongsup S, Scorza V (2010) Update on the diagnosis and management of Giardia spp. infections in dogs and cats. Top Companion Anim Med 25:155–162. https://doi.org/10.1053/j.tcam.2010.07.003
Tarsitano E, Greco G, Decaro N et al (2010) Environmental monitoring and analysis of faecal contamination in an urban setting in the city of Bari (Apulia region, Italy): health and hygiene implications. Int J Environ Res Public Health 7:3972–3986. https://doi.org/10.3390/ijerph7113972
Thompson RCA (2004) The zoonotic significance and molecular epidemiology of Giardia and giardiasis. Vet Parasitol 126:15–35. https://doi.org/10.1016/j.vetpar.2004.09.008
Traversa D, Frangipane di Regalbono A, Di Cesare A et al (2014) Environmental contamination by canine geohelminths. Parasit Vectors 7:67. https://doi.org/10.1186/1756-3305-7-67
Uehlinger FD, Greenwood SJ, McClure JT et al (2013) Zoonotic potential of Giardia duodenalis and Cryptosporidium spp. and prevalence of intestinal parasites in young dogs from different populations on Prince Edward Island, Canada. Vet Parasitol 196:509–514. https://doi.org/10.1016/j.vetpar.2013.03.020
Upjohn M, Cobb C, Monger J et al (2010) Prevalence, molecular typing and risk factor analysis for Giardia duodenalis infections in dogs in a central London rescue shelter. Vet Parasitol 172:341–346. https://doi.org/10.1016/j.vetpar.2010.05.010
Verweij JJ, Schinkel J, Laeijendecker D et al (2003) Real-time PCR for the detection of Giardia lamblia. Mol Cell Probes 17:223–225. https://doi.org/10.1016/S0890-8508(03)00057-4
Wang A, Ruch-Gallie R, Scorza V et al (2012) Prevalence of Giardia and Cryptosporidium species in dog park attending dogs compared to non-dog park attending dogs in one region of Colorado. Vet Parasitol 184:335–340. https://doi.org/10.1016/j.vetpar.2011.08.019
Xiao L, Bern C, Limor J et al (2001) Identification of 5 types of Cryptosporidium parasites in children in Lima, Peru. J Infect Dis 183:492–497. https://doi.org/10.1086/318090
Xiao L, Fayer R, Ryan U, Upton SJ (2004) Cryptosporidium taxonomy: recent advances and implications for public health. Clin Microbiol Rev 17:72–97. https://doi.org/10.1128/CMR.17.1.72
Yoshiuchi R, Matsubayashi M, Kimata I et al (2010) Survey and molecular characterization of Cryptosporidium and Giardia spp. in owned companion animal, dogs and cats, in Japan. Vet Parasitol 174:313–316. https://doi.org/10.1016/j.vetpar.2010.09.004
Zanzani SAS, Gazzonis AL, Scarpa P et al (2014) Intestinal parasites of owned dogs and cats from metropolitan and micropolitan areas: prevalence, zoonotic risks, and pet owner awareness in Northern Italy. Biomed Res Int 2014:1–10. https://doi.org/10.1155/2014/696508
Acknowledgments
This work was supported by the research grant of the University of Padua Prot. CPDA110843. MP, AFdR and DT conceived the study and all authors contributed to its drafting, preparation and intellectual content. MP and AFdR were scientifically responsible for the studies whose unpublished data are reported. All authors read and approved the final manuscript.
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Simonato, G., Frangipane di Regalbono, A., Cassini, R. et al. Molecular detection of Giardia duodenalis and Cryptosporidium spp. in canine faecal samples contaminating public areas in Northern Italy. Parasitol Res 116, 3411–3418 (2017). https://doi.org/10.1007/s00436-017-5671-z
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DOI: https://doi.org/10.1007/s00436-017-5671-z
Keywords
- Protozoa
- Giardia duodenalis
- Cryptosporidium
- Environmental faecal pollution
- Public health
- Italy