Molecular characterization of Cryptosporidium isolates from high-excreting young dairy calves in dairy cattle herds in Western France

Abstract

Ninety-two Cryptosporidium sp.-positive fecal samples of dairy diarrheic or non-diarrheic calves from 30 cattle herds in Normandy (France) were selected. Here, the aim was to investigate the species of Cryptosporidium excreted as well as the subtypes of Cryptosporidium parvum found in 7-17-day-old dairy calves. Excretion levels were comprised between 2 × 10(4) and 4 × 10(7) oocysts per gram of feces. Here, a nested 18S SSU rRNA PCR associated with sequencing was performed for identification of Cryptosporidium species and revealed the presence of C. parvum in most cases (80/82), except for two animals which were infected with Cryptosporidium bovis. Then, C. parvum samples were submitted to gp60 PCR. For 39 samples from 24 different herds, a multilocus analysis based on four mini-microsatellites loci (MM19, MM5, MSF, and MS9-Mallon) were conducted. These results were combined with sequence analysis of the gp60 to obtain multilocus types (MLTs). Here, C. parvum gp60 genotyping identified three subtypes in the IIa zoonotic allele family: IIaA15G2R1 (88%), IIaA16G3R1 (10%), and IIaA19G2R1 (2%), and we identified 12 MLTs. The MS9-Mallon locus was reported as the most polymorphic (five alleles). The most common MLT was MLT 1 with 15 samples in 10 farms: (MS9-M: 298, MSF: 165, MM5: 264, MM19: 462, and gp60 subtype: IIaA15G2R1). When comparing diarrheic and non-diarrheic fecal samples, no difference was seen for distribution of Cryptosporidium species, C. parvum gp60 subtypes, and MLTs. Here, in a range of oocyst excretion of 10(4)-10(7) opg, both in diarrheic and non-diarrheic calves, infection was mainly due to C. parvum and to the zoonotic subtype: IIaA15G2R1.

Genetic characterization of Cryptosporidium in animal and human isolates from Jordan

“…However, the IIaA19G2R1 subtype is a relatively rare subtype and has previously been described by Xiao et al (2007), Wielinga et al (2008), Ng et al (2012), Rieux et al (2013) and Couto et al (2014) infecting cattle in the North America, The Netherlands, Australia, France and Brazil, respectively and by Nolan et al (2013) in deer in Australia. To the best of our knowledge, this is the first report of this subtype in sheep.…”

Molecular characterization of Cryptosporidium isolates from diarrheal dairy calves in France

“…Thus far, only a few studies have used molecular tools to characterise Cryptosporidium spp. in cattle in 15 of the 95 metropolitan French departments (administrative regions): Allier, Côtes-d'Armor, Ille-et-Vilaine, Landes, Mayenne, Morbihan, Moselle, Orne, Pas-de-Calais, Puy-de-Dôme, Pyrénées-Atlantiques, Hautes-Pyrénées, Deux-Sèvres, Tarn, and Vendée (Follet et al, 2011;Ngouanesavanh et al, 2006;Rieux et al, 2014Rieux et al, , 2013bRieux et al, , 2013cRieux et al, , 2013a. Little is known about which Cryptosporidium spp.…”

“…infects calves in other French departments. However, subtype analysis solely on the basis of C. parvum gp60 amplicons is insufficient (Follet et al, 2011;Rieux et al, 2014Rieux et al, , 2013cRieux et al, , 2013a, thus there is a strong need for more molecular epidemiological data on French bovine cryptosporidiosis to fully determine the breadth of C. parvum genetic diversity. Thus, the aim of this study was to characterise Cryptosporidium isolates in calves from different French departments: Allier, Ardèche, Côte-d'Or, Moselle, Saôneet-Loire, and Yonne.…”

Surto de criptosporidiose em bezerros no Sul do Rio Grande do Sul

“…Criptosporidiose é uma enfermidade de ocorrência mundial causada por protozoários do gênero Cryptosporidium, que infectam uma ampla gama de hospedeiros (mamíferos, aves, répteis, anfíbios e peixes), inclusive o homem (Rieux et al 2013 This paper describes the epidemiology, clinical signs and pathology of an outbreak of cryptosporidiosis in calves in Southern Brazil. Thirty-five out of 400 calves with 30-45 days of age were affected and 16 died.…”