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Integrated epidemiological and molecular analysis of Cryptosporidium spp. and Giardia duodenalis isolates in dairy calves from Terceira Island, Azores.

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Τίτλος: Integrated epidemiological and molecular analysis of Cryptosporidium spp. and Giardia duodenalis isolates in dairy calves from Terceira Island, Azores.
Συγγραφείς: Louro M; CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Avenida da Universidade Técnica, Lisbon, 1300-477, Portugal.; Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Lisbon, Portugal.; Laboratório de Parasitologia, Instituto Nacional de Investigação Agrária e Veterinária (INIAV), Oeiras, 2780-157, Portugal., Linhares JCT; Institute of Agricultural and Environmental Research and Technology (IITAA), University of the Azores, Angra do Heroísmo, 9500-321, Portugal.; UNICOL - Cooperativa Agrícola, C.R.L, Angra do Heroísmo, Portugal., Pinto CA; CBA - Biotechnology Centre of Azores, University of the Azores, Rua Capitão João d'Ávila, Angra do Heroísmo, 9700-042, Portugal., Pino-Vera R; Instituto Universitario de Enfermedades Tropicales y Salud Pública de Canarias, Universidad de La Laguna (ULL), Av. Astrofísico F. Sánchez, sn, La Laguna, Canary Islands, 38203, Spain.; Departamento de Obstetricia y Ginecología, Pediatría, Medicina Preventiva y Salud Pública, Toxicología, Medicina Legal y Forense y Parasitología, Facultad de Farmacia, Universidad de La Laguna (ULL), La Laguna, Canary Islands, Spain.; Programa de Doctorado de Ciencias Médicas y Farmacéuticas, Desarrollo y Calidad de Vida, Universidad de La Laguna (ULL), La Laguna, Spain., Nunes T; CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Avenida da Universidade Técnica, Lisbon, 1300-477, Portugal.; Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Lisbon, Portugal., Gomes J; CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Avenida da Universidade Técnica, Lisbon, 1300-477, Portugal. jacinto.gomes@ipportalegre.pt.; Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Lisbon, Portugal. jacinto.gomes@ipportalegre.pt.; Elvas School of Biosciences, Polytechnic Institute of Portalegre, Portalegre, Portugal. jacinto.gomes@ipportalegre.pt.; VALORIZA - Research Centre for Endogenous Resources Valorization, Polytechnic Institute of Portalegre, Portalegre, Portugal. jacinto.gomes@ipportalegre.pt., Pereira da Fonseca I; CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Avenida da Universidade Técnica, Lisbon, 1300-477, Portugal.; Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Lisbon, Portugal.
Πηγή: Parasitology research [Parasitol Res] 2025 Dec 05; Vol. 124 (12), pp. 151. Date of Electronic Publication: 2025 Dec 05.
Τύπος έκδοσης: Journal Article
Γλώσσα: English
Στοιχεία περιοδικού: Publisher: Springer International Country of Publication: Germany NLM ID: 8703571 Publication Model: Electronic Cited Medium: Internet ISSN: 1432-1955 (Electronic) Linking ISSN: 09320113 NLM ISO Abbreviation: Parasitol Res Subsets: MEDLINE
Imprint Name(s): Original Publication: Berlin : Springer International, c1987-
Ιατρικοί όροι (MeSH): Cryptosporidiosis*/epidemiology , Cryptosporidiosis*/parasitology , Cryptosporidium*/genetics , Cryptosporidium*/isolation & purification , Cryptosporidium*/classification , Giardiasis*/epidemiology , Giardiasis*/veterinary , Giardiasis*/parasitology , Cattle Diseases*/epidemiology , Cattle Diseases*/parasitology , Giardia lamblia*/genetics , Giardia lamblia*/isolation & purification , Giardia lamblia*/classification, Feces/parasitology ; Azores/epidemiology ; DNA, Protozoan/genetics ; Animals ; Cattle ; Prevalence ; Genotype ; Genetic Variation ; Risk Factors
Περίληψη: Cryptosporidium spp. and Giardia duodenalis are major enteric pathogens in neonatal calves, contributing to gastrointestinal disease and raising concerns due to their zoonotic potential. Despite their global distribution, little is known about their occurrence and genetic diversity in insular regions such as the Azores archipelago (Portugal). This study investigated the prevalence, distribution, genetic diversity, and farm-level risk factors associated with these parasites in dairy calves from Terceira Island. Fecal samples (n = 142) from 28 farms were examined by microscopy, direct immunofluorescence assay (DFA), and PCR targeting multiple loci (SSU rRNA, gp60, LSU rRNA for Cryptosporidium spp.; gdh, bg, tpi for G. duodenalis). Sequencing enabled molecular identification of species, genotypes, and assemblages. Farm management data were collected by standardized questionnaires, and spatial analysis used farm location. Overall, 65.5% (93/142) of calves were infected with at least one parasite, with Cryptosporidium spp. and Giardia spp. detected in 42.3% (60/142) and 44.4% (63/142) of samples, respectively. Mixed infections occurred in 21.1% (30/142) of calves. Molecular characterization identified C. parvum (subtype IIaA15G2R1), C. bovis, and C. ryanae, as well as G. duodenalis assemblage E (predominant) and assemblage A (rare). Spatial mapping showed Cryptosporidium spp. to be widespread, while Giardia spp. displayed localized clusters. Use of paromomycin and higher hygiene scores were associated with reduced Giardia risk, whereas no risk factors were found for Cryptosporidium. These findings provide novel insights into protozoan infections in calves in the Azores and highlight the value of integrating molecular, spatial, and management data for targeted control strategies.
(© 2025. The Author(s).)
Competing Interests: Declarations. Ethical approval: The study was reviewed and approved by the Ethics Committee for Research and Teaching (CEIE) of the Faculty of Veterinary Medicine, University of Lisbon (CEIE N/Ref. 039/2023). Farmers provided written informed consent for the participation of their calves. Competing interests: The authors declare no competing interests. Clinical trial number: Not applicable.
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Exp Parasitol. 2010 Jan;124(1):80-9. (PMID: 19358845)
Appl Environ Microbiol. 2000 Dec;66(12):5492-8. (PMID: 11097935)
J Clin Microbiol. 2003 Jun;41(6):2744-7. (PMID: 12791920)
Pathogens. 2022 Jan 15;11(1):. (PMID: 35056051)
Vet Clin North Am Food Anim Pract. 2020 Mar;36(1):223-238. (PMID: 32029186)
J Dairy Sci. 2018 Oct;101(10):9199-9213. (PMID: 29859689)
Acta Vet Scand. 2003;44(3-4):145-52. (PMID: 15074627)
Int J Parasitol. 2005 Feb;35(2):207-13. (PMID: 15710441)
Parasit Vectors. 2017 Oct 23;10(1):509. (PMID: 29061171)
Parasit Vectors. 2019 Mar 12;12(1):95. (PMID: 30867023)
J Vet Med B Infect Dis Vet Public Health. 2006 Jun;53(5):244-6. (PMID: 16732884)
Emerg Infect Dis. 2003 Nov;9(11):1444-52. (PMID: 14718089)
Vet Parasitol. 2006 Sep 10;140(3-4):204-16. (PMID: 16647210)
Infect Genet Evol. 2004 Jun;4(2):125-30. (PMID: 15157630)
Nucleic Acids Res. 2016 Jul 8;44(W1):W232-5. (PMID: 27084950)
Curr Res Parasitol Vector Borne Dis. 2025 May 08;7:100264. (PMID: 40487329)
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Grant Information: UI/BD/152818/2022 Fundação para a Ciência e a Tecnologia; Tesis 2022010038 European Social Fund Plus
Contributed Indexing: Keywords: Dairy calves; Molecular epidemiology; Portugal.; Prevalence; Risk factors; Zoonoses
Substance Nomenclature: 0 (DNA, Protozoan)
Entry Date(s): Date Created: 20251205 Date Completed: 20251206 Latest Revision: 20260919
Update Code: 20260919
PubMed Central ID: PMC12680760
DOI: 10.1007/s00436-025-08613-x
PMID: 41350959
Βάση Δεδομένων: MEDLINE
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  Label: Title
  Group: Ti
  Data: Integrated epidemiological and molecular analysis of Cryptosporidium spp. and Giardia duodenalis isolates in dairy calves from Terceira Island, Azores.
– Name: Author
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  Data: <searchLink fieldCode="AU" term="%22Louro+M%22">Louro M</searchLink>; CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Avenida da Universidade Técnica, Lisbon, 1300-477, Portugal.; Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Lisbon, Portugal.; Laboratório de Parasitologia, Instituto Nacional de Investigação Agrária e Veterinária (INIAV), Oeiras, 2780-157, Portugal.<br /><searchLink fieldCode="AU" term="%22Linhares+JCT%22">Linhares JCT</searchLink>; Institute of Agricultural and Environmental Research and Technology (IITAA), University of the Azores, Angra do Heroísmo, 9500-321, Portugal.; UNICOL - Cooperativa Agrícola, C.R.L, Angra do Heroísmo, Portugal.<br /><searchLink fieldCode="AU" term="%22Pinto+CA%22">Pinto CA</searchLink>; CBA - Biotechnology Centre of Azores, University of the Azores, Rua Capitão João d'Ávila, Angra do Heroísmo, 9700-042, Portugal.<br /><searchLink fieldCode="AU" term="%22Pino-Vera+R%22">Pino-Vera R</searchLink>; Instituto Universitario de Enfermedades Tropicales y Salud Pública de Canarias, Universidad de La Laguna (ULL), Av. Astrofísico F. Sánchez, sn, La Laguna, Canary Islands, 38203, Spain.; Departamento de Obstetricia y Ginecología, Pediatría, Medicina Preventiva y Salud Pública, Toxicología, Medicina Legal y Forense y Parasitología, Facultad de Farmacia, Universidad de La Laguna (ULL), La Laguna, Canary Islands, Spain.; Programa de Doctorado de Ciencias Médicas y Farmacéuticas, Desarrollo y Calidad de Vida, Universidad de La Laguna (ULL), La Laguna, Spain.<br /><searchLink fieldCode="AU" term="%22Nunes+T%22">Nunes T</searchLink>; CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Avenida da Universidade Técnica, Lisbon, 1300-477, Portugal.; Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Lisbon, Portugal.<br /><searchLink fieldCode="AU" term="%22Gomes+J%22">Gomes J</searchLink>; CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Avenida da Universidade Técnica, Lisbon, 1300-477, Portugal. jacinto.gomes@ipportalegre.pt.; Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Lisbon, Portugal. jacinto.gomes@ipportalegre.pt.; Elvas School of Biosciences, Polytechnic Institute of Portalegre, Portalegre, Portugal. jacinto.gomes@ipportalegre.pt.; VALORIZA - Research Centre for Endogenous Resources Valorization, Polytechnic Institute of Portalegre, Portalegre, Portugal. jacinto.gomes@ipportalegre.pt.<br /><searchLink fieldCode="AU" term="%22Pereira+da+Fonseca+I%22">Pereira da Fonseca I</searchLink>; CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Avenida da Universidade Técnica, Lisbon, 1300-477, Portugal.; Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Lisbon, Portugal.
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  Data: <searchLink fieldCode="JN" term="%228703571%22">Parasitology research</searchLink> [Parasitol Res] 2025 Dec 05; Vol. 124 (12), pp. 151. <i>Date of Electronic Publication: </i>2025 Dec 05.
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  Data: <i>Publisher: </i><searchLink fieldCode="PB" term="%22Springer+International%22">Springer International </searchLink><i>Country of Publication: </i>Germany <i>NLM ID: </i>8703571 <i>Publication Model: </i>Electronic <i>Cited Medium: </i>Internet <i>ISSN: </i>1432-1955 (Electronic) <i>Linking ISSN: </i><searchLink fieldCode="IS" term="%2209320113%22">09320113 </searchLink><i>NLM ISO Abbreviation: </i>Parasitol Res <i>Subsets: </i>MEDLINE
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  Data: <i>Original Publication</i>: Berlin : Springer International, c1987-
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  Data: <searchLink fieldCode="MM" term="%22Cryptosporidiosis%22">Cryptosporidiosis*</searchLink>/<searchLink fieldCode="MM" term="%22Cryptosporidiosis+epidemiology%22">epidemiology</searchLink> <br /><searchLink fieldCode="MM" term="%22Cryptosporidiosis%22">Cryptosporidiosis*</searchLink>/<searchLink fieldCode="MM" term="%22Cryptosporidiosis+parasitology%22">parasitology</searchLink> <br /><searchLink fieldCode="MM" term="%22Cryptosporidium%22">Cryptosporidium*</searchLink>/<searchLink fieldCode="MM" term="%22Cryptosporidium+genetics%22">genetics</searchLink> <br /><searchLink fieldCode="MM" term="%22Cryptosporidium%22">Cryptosporidium*</searchLink>/<searchLink fieldCode="MM" term="%22Cryptosporidium+isolation+%26+purification%22">isolation & purification</searchLink> <br /><searchLink fieldCode="MM" term="%22Cryptosporidium%22">Cryptosporidium*</searchLink>/<searchLink fieldCode="MM" term="%22Cryptosporidium+classification%22">classification</searchLink> <br /><searchLink fieldCode="MM" term="%22Giardiasis%22">Giardiasis*</searchLink>/<searchLink fieldCode="MM" term="%22Giardiasis+epidemiology%22">epidemiology</searchLink> <br /><searchLink fieldCode="MM" term="%22Giardiasis%22">Giardiasis*</searchLink>/<searchLink fieldCode="MM" term="%22Giardiasis+veterinary%22">veterinary</searchLink> <br /><searchLink fieldCode="MM" term="%22Giardiasis%22">Giardiasis*</searchLink>/<searchLink fieldCode="MM" term="%22Giardiasis+parasitology%22">parasitology</searchLink> <br /><searchLink fieldCode="MM" term="%22Cattle+Diseases%22">Cattle Diseases*</searchLink>/<searchLink fieldCode="MM" term="%22Cattle+Diseases+epidemiology%22">epidemiology</searchLink> <br /><searchLink fieldCode="MM" term="%22Cattle+Diseases%22">Cattle Diseases*</searchLink>/<searchLink fieldCode="MM" term="%22Cattle+Diseases+parasitology%22">parasitology</searchLink> <br /><searchLink fieldCode="MM" term="%22Giardia+lamblia%22">Giardia lamblia*</searchLink>/<searchLink fieldCode="MM" term="%22Giardia+lamblia+genetics%22">genetics</searchLink> <br /><searchLink fieldCode="MM" term="%22Giardia+lamblia%22">Giardia lamblia*</searchLink>/<searchLink fieldCode="MM" term="%22Giardia+lamblia+isolation+%26+purification%22">isolation & purification</searchLink> <br /><searchLink fieldCode="MM" term="%22Giardia+lamblia%22">Giardia lamblia*</searchLink>/<searchLink fieldCode="MM" term="%22Giardia+lamblia+classification%22">classification</searchLink><br /><searchLink fieldCode="MH" term="%22Feces%22">Feces</searchLink>/<searchLink fieldCode="MH" term="%22Feces+parasitology%22">parasitology</searchLink> ; <searchLink fieldCode="MH" term="%22Azores%22">Azores</searchLink>/<searchLink fieldCode="MH" term="%22Azores+epidemiology%22">epidemiology</searchLink> ; <searchLink fieldCode="MH" term="%22DNA%2C+Protozoan%22">DNA, Protozoan</searchLink>/<searchLink fieldCode="MH" term="%22DNA%2C+Protozoan+genetics%22">genetics</searchLink> ; <searchLink fieldCode="MH" term="%22Animals%22">Animals</searchLink> ; <searchLink fieldCode="MH" term="%22Cattle%22">Cattle</searchLink> ; <searchLink fieldCode="MH" term="%22Prevalence%22">Prevalence</searchLink> ; <searchLink fieldCode="MH" term="%22Genotype%22">Genotype</searchLink> ; <searchLink fieldCode="MH" term="%22Genetic+Variation%22">Genetic Variation</searchLink> ; <searchLink fieldCode="MH" term="%22Risk+Factors%22">Risk Factors</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: Cryptosporidium spp. and Giardia duodenalis are major enteric pathogens in neonatal calves, contributing to gastrointestinal disease and raising concerns due to their zoonotic potential. Despite their global distribution, little is known about their occurrence and genetic diversity in insular regions such as the Azores archipelago (Portugal). This study investigated the prevalence, distribution, genetic diversity, and farm-level risk factors associated with these parasites in dairy calves from Terceira Island. Fecal samples (n = 142) from 28 farms were examined by microscopy, direct immunofluorescence assay (DFA), and PCR targeting multiple loci (SSU rRNA, gp60, LSU rRNA for Cryptosporidium spp.; gdh, bg, tpi for G. duodenalis). Sequencing enabled molecular identification of species, genotypes, and assemblages. Farm management data were collected by standardized questionnaires, and spatial analysis used farm location. Overall, 65.5% (93/142) of calves were infected with at least one parasite, with Cryptosporidium spp. and Giardia spp. detected in 42.3% (60/142) and 44.4% (63/142) of samples, respectively. Mixed infections occurred in 21.1% (30/142) of calves. Molecular characterization identified C. parvum (subtype IIaA15G2R1), C. bovis, and C. ryanae, as well as G. duodenalis assemblage E (predominant) and assemblage A (rare). Spatial mapping showed Cryptosporidium spp. to be widespread, while Giardia spp. displayed localized clusters. Use of paromomycin and higher hygiene scores were associated with reduced Giardia risk, whereas no risk factors were found for Cryptosporidium. These findings provide novel insights into protozoan infections in calves in the Azores and highlight the value of integrating molecular, spatial, and management data for targeted control strategies.<br /> (© 2025. The Author(s).)
– Name: Abstract
  Label: Competing Interests
  Group: Ab
  Data: Declarations. Ethical approval: The study was reviewed and approved by the Ethics Committee for Research and Teaching (CEIE) of the Faculty of Veterinary Medicine, University of Lisbon (CEIE N/Ref. 039/2023). Farmers provided written informed consent for the participation of their calves. Competing interests: The authors declare no competing interests. Clinical trial number: Not applicable.
– Name: Ref
  Label: References
  Group: RefInfo
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  Data: UI/BD/152818/2022 Fundação para a Ciência e a Tecnologia; Tesis 2022010038 European Social Fund Plus
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  Data: <i>Keywords: </i>Dairy calves; Molecular epidemiology; Portugal.; Prevalence; Risk factors; Zoonoses
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        Value: 10.1007/s00436-025-08613-x
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        Text: English
    PhysicalDescription:
      Pagination:
        StartPage: 151
    Subjects:
      – SubjectFull: Feces parasitology
        Type: general
      – SubjectFull: Azores epidemiology
        Type: general
      – SubjectFull: DNA, Protozoan genetics
        Type: general
      – SubjectFull: Animals
        Type: general
      – SubjectFull: Cattle
        Type: general
      – SubjectFull: Prevalence
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      – SubjectFull: Genotype
        Type: general
      – SubjectFull: Genetic Variation
        Type: general
      – SubjectFull: Risk Factors
        Type: general
      – SubjectFull: Cryptosporidiosis epidemiology
        Type: general
      – SubjectFull: Cryptosporidiosis parasitology
        Type: general
      – SubjectFull: Cryptosporidium genetics
        Type: general
      – SubjectFull: Cryptosporidium isolation & purification
        Type: general
      – SubjectFull: Cryptosporidium classification
        Type: general
      – SubjectFull: Giardiasis epidemiology
        Type: general
      – SubjectFull: Giardiasis veterinary
        Type: general
      – SubjectFull: Giardiasis parasitology
        Type: general
      – SubjectFull: Cattle Diseases epidemiology
        Type: general
      – SubjectFull: Cattle Diseases parasitology
        Type: general
      – SubjectFull: Giardia lamblia genetics
        Type: general
      – SubjectFull: Giardia lamblia isolation & purification
        Type: general
      – SubjectFull: Giardia lamblia classification
        Type: general
    Titles:
      – TitleFull: Integrated epidemiological and molecular analysis of Cryptosporidium spp. and Giardia duodenalis isolates in dairy calves from Terceira Island, Azores.
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              M: 12
              Text: 2025 Dec 05
              Type: published
              Y: 2025
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            – TitleFull: Parasitology research
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