Genetic uniqueness of Cryptosporidium parvum from dairy calves in Colombia

Avendaño, Catalina; Ramo, Ana; Vergara-Castiblanco, Claudia; Sánchez-Acedo, Caridad; Quílez, Joaquín

doi:10.1007/s00436-018-5818-6

Cited by 28 publications

(27 citation statements)

References 60 publications

(55 reference statements)

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“…At least one C. parvum-positive calf was detected in 89% of the herds studied. Highly variable herd prevalence ranging from 50.5 to 96% have been reported (Hamnes et al 2006;Brook et al 2008;Trotz-Williams et al 2008;Silverlås et al 2009;Tiranti et al 2011;Smith et al 2014;Al Mawly et al 2015;Delafosse et al 2015;Garro et al 2016;Avendaño et al 2018). Similar to the considerable differences of overall prevalence that have been reported, corresponding dissimilarities in herd prevalence may likewise be attributed to the different sensitivities of applied diagnostic test.…”

Section: Dairy Herdmentioning

confidence: 69%

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Prevalence of Cryptosporidium parvum in dairy calves and GP60 subtyping of diarrheic calves in central Argentina

et al. 2019

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Cryptosporidiosis of calves is caused by the enteroprotozoan Cryptosporidium spp. The disease results in intense diarrhea of calves associated with substantial economic losses in dairy farming worldwide. The aim of this study was to determine calf, herd, and within-herd Cryptosporidium prevalence and identify Cryptosporidium species and subtypes in calves with diarrhea in intensive dairy herds in central Argentina. A total of 1073 fecal samples were collected from 54 randomly selected dairy herds. Cryptosporidium-oocysts were isolated and concentrated from fecal samples using formol-ether and detected by light microscopy with the modified Ziehl-Neelsen technique. Overall prevalence of oocyst-excreting calves was found to be 25.5% (274/ 1073) (95% C.I. 22.9; 28.1%). Of the herds studied, 89% (48/54) included at least one infected calf, whereas within-herd prevalence ranged from the absence of infection to 57% (20/35). A highly significant association was found between the presence of diarrhea and C. parvum infection (χ 2 = 55.89, p < 0.001). For species determination, genomic DNA isolated from oocystpositive fecal samples was subjected to PCR-RFLP of the 18S rRNA gene resulting exclusively in Cryptosporidium parvum identification. C. parvum isolates of calves displaying diarrhea and high rate of excretion of oocysts were subtyped by PCR amplification and direct sequencing of the 60 kDa glycoprotein (GP60) gene. Altogether five GP60 subtypes, designated IIaA18G1R1, IIaA20G1R1, IIaA21G1R1, IIaA22G1R1, and IIaA24G1R1 were identified. Interestingly, IIaA18G1R1 and IIaA20G1R1 were predominant in calves with diarrhea and high infection intensity. Notably, IIaA24G1R1 represents a novel, previously unrecognized C. parvum subtype. The subtype IIaA18G1R1, frequently found in this study, is strongly implicated in zoonotic transmission. These results suggest that calves might be an important source for human cryptosporidiosis in Argentina.

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Section: Dairy Herdmentioning

confidence: 69%

“…reported that cryptosporidiosis risk is higher in calves younger than 30 days old (Tiranti et al 2011;Smith et al 2014;Delafosse et al 2015;Avendaño et al 2018).…”

Section: Introductionmentioning

confidence: 98%

Prevalence of Cryptosporidium parvum in dairy calves and GP60 subtyping of diarrheic calves in central Argentina

et al. 2019

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“…A wide range of subtypes belonging to a single gp60 allelic family have been reported in Brazil (IIaA14G2R1, IIaA14G2R2, IIaA15G2R1, IIaA16G3R2, IIaA17G2R1, IIaA18G1R1, IIaA18G2R2, IIaA19G2R1, IIaA19G2R2, IIaA20G1R1; IIaA20G2R1, IIaA20G2R2) (Meireles et al, 2011;Paz e Silva et al, 2013;do Couto et al, 2014;Heckler et al, 2015;Toledo et al, 2017), Argentina (IIaA16G1R1, IIaA17G1R1, IIaA18G1R1, IIaA19G1R1, IIaA20G1R1, IIaA21G1R1, IIaA22G1R1, IIaA23G1R1) (Tomazic et al, 2013;Del Coco et al, 2014) and Chile (IIaA15G4R1, IIaA16G4R1, IIaA17G4R1) (Mercado et al, 2015). The fact that many of these subtypes were novel allelic variants or were not shared among different countries has been related to the geographic isolation of the protozoan in cattle farms (Avendaño et al, 2018). The latter authors conducted the first large study of Cryptosporidium species and subtypes in Colombia, which revealed the distinct identity of C. parvum circulating in calves.…”

Section: Discussionmentioning

confidence: 99%

“…Genomic DNA of 70 C. parvum isolates from a previous study was used in this molecular analysis (Avendaño et al, 2018). These isolates were collected between 2010 and 2012 from naturally infected diarrhoeic (n: 25) and non-diarrhoeic (n: 45) calves younger than 35 days.…”

Section: Cryptosporidium Isolatesmentioning

confidence: 99%

“…Modest numbers of specimens from calves have been characterised by gp60 sequencing in Brazil, Argentina or Chile (Meireles et al, 2011;Paz e Silva et al, 2013;Tomazic et al, 2013;Del Coco et al, 2014;do Couto et al, 2014;Heckler et al, 2015;Mercado et al, 2015;Toledo et al, 2017) and no previous work with VNTR markers has so far been conducted. In Colombia, the single genotyping study reporting the distribution of Cryptosporidium species and gp60 subtypes among pre-weaned calves highlighted this protozoan as a common and widespread pathogen in the central area of the country (Avendaño et al, 2018). Moreover, the latter study revealed the genetic distinctiveness of C. parvum in this geographical area, with the presence of eight subtypes within the IIa family, but the overwhelming predominance of an unusual subtype (IIaA18G5R1) previously unreported in natural infections in human or animal hosts.…”

Section: Introductionmentioning

confidence: 99%

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Multilocus fragment analysis of Cryptosporidium parvum from pre-weaned calves in Colombia

et al. 2019

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A B S T R A C TThe intra-species genetic diversity of Cryptosporidium parvum in dairy cattle farms in the central area of Colombia was investigated using a multilocus fragment typing approach with nine variable-number tandem-repeat (VNTR) loci and the gp60 gene. Genomic DNA of 70 C. parvum isolates from pre-weaned calves in 32 farms was analysed. Most markers showed two (ML1, MSB, CP47, and MSC6-7) or three alleles (5B12, Cgd2_3850, and Cgd6_5400), although they exhibited a major allele accounting for more than 69% of specimens, which explains their low discriminatory index. The TP14 microsatellite was monomorphic while a total of six alleles were found at the ML2 microsatellite. The two novel allelic variants (219bp, 245bp) exhibited by more than 36% of specimens at the latter locus were a remarkable finding. The 10-markers typing tool provided a Hunter-Gaston discriminatory value of 0.940 (95% CI, 0.918 -0.961) and differentiated 22 multilocus subtypes (MLTs). Nevertheless, the combination of the three most informative markers (ML2, gp60, and Cgd2_3850) differentiated 68% of MLTs and hardly impaired the discriminatory index. The fact that many MLTs (13/22) were distinctive for individual farms provides evidence for the endemic nature of the infection and the major role played by transmission within farms. The eBURST algorithm suggested a low degree of genetic divergence. All but three MLTs were clustered in a clonal complex with a star-like topology typical of clonal expansion, however linkage analysis did not find evidence of linkage disequilibrium. Bayesian analysis also identified a genetic structure with K = 3 being the best estimation of ancestral clusters, although a large proportion of isolates (35%) could not be allocated to a single population, which indicates their mixed origin. The results confirm the genetic distinctiveness of C. parvum in cattle farms in this geographical area. This is the first multilocus analysis on the intra-specific variability of Cryptosporidium from calves in South America.

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Molecular evaluation of Cryptosporidium spp. among breeding calves of Lorestan province Western Iran

Talebi

Mamaghani

Kheirandish

et al. 2022

Veterinary Medicine & Sci

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Background Cryptosporidium spp. are opportunistic intestinal protozoans with global distribution and are of great importance as zoonotic protozoans are common to humans and domestic animals, including cattle and calves. Identification and detection of parasite species using precise methods including molecular methods can be an effective step in treating and controlling parasites. Objectives This study aimed to investigate the prevalence of Cryptosporidium among breeding calves of Khorramabad city, Lorestan province, Western Iran, using PCR. Methods The faecal samples were taken from 181 healthy and diarrhoeal calves and after the Ziehl Neelsen Acid‐fast staining and microscopic evaluation, the genomic DNA was extracted for molecular evaluations. To detect Cryptosporidium species, specific primers targeting the SAM‐1 gene of Cryptosporidium and a commercial master mix were used for PCR. Results Out of 181 faecal samples of breeding calves in Khorramabad city, 9 samples (5%) were positive for Cryptosporidium spp. using the PCR method. Statistical analysis of the data showed that there was no significant statistical relationship between Cryptosporidium infection of the calves and variables of age, breed, type of water consumption, clinical signs of diarrhoea, and sampling location, while parasite infection had a significant relationship with calf gender so that all Cryptosporidium positive samples were from male calves ( p ≤ 0.05). Conclusions Considering the presence of Cryptosporidium infection, the region's traditional grazing system, and the close relationship between livestock and humans, there is a possibility of human infection in the region. So preventive measures such as periodic animal testing with sensitive and accurate diagnostic techniques including PCR, pharmacological treatment of livestock, water hygiene and the use of industrial grazing instead of traditional grazing to improve the hygiene of food consumed by livestock are recommended.

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Genetic uniqueness of Cryptosporidium parvum from dairy calves in Colombia

Cited by 28 publications

References 60 publications

Prevalence of Cryptosporidium parvum in dairy calves and GP60 subtyping of diarrheic calves in central Argentina

Prevalence of Cryptosporidium parvum in dairy calves and GP60 subtyping of diarrheic calves in central Argentina

Multilocus fragment analysis of Cryptosporidium parvum from pre-weaned calves in Colombia

Molecular evaluation of Cryptosporidium spp. among breeding calves of Lorestan province Western Iran

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