Identification of Cryptosporidiumspecies and genotypes in dairy cattle in Brazil

Abstract

In this study, we identified Cryptosporidium species and genotypes present in dairy cattle in the central region of São Paulo state, Brazil. Fecal specimens were collected from 200 animals (100 calves and 100 cows) in ten dairy farms. Fecal samples were examined using microscopic examination (ME), enzyme immunoassay (EIA) and polymerase chain reaction (PCR). Cryptosporidium species and genotypes were determined by restriction fragment length polymorphism (RFLP) or DNA sequencing analysis of the SSU-rRNA and GP60 genes. The occurrence of Cryptosporidium spp. infection was 14% (28/200). The occurrence in calves (26%) was significantly higher than in cows (2%). Of the 27 Cryptosporidium-positive specimens submitted to genotyping, C. andersoni was identified in 23 (85.1%), C. bovis in three (11.1%), and the zoonotic C. parvum subtype IIaA15G2R1 in one (3.7%). The study demonstrates that Cryptosporidium spp. infection was common and widespread in dairy cattle in this region and that calves have a high prevalence of C. andersoni. Furthermore, the presence of C. parvum subtype IIaA15G2R1 indicates that dairy calves from this region should be considered a potential source of zoonotic Cryptosporidium oocysts.Keywords: Cryptosporidium spp., cattle, PCR-RFLP, sequencing, genotyping. ResumoNo presente estudo foram identificadas espécies e genótipos de Cryptosporidium originadas de bovinos leiteiros na região central do estado de São Paulo, Brasil. Amostras fecais foram coletadas de 200 animais (100 bezerros e 100 vacas) em 10 propriedades leiteiras. As amostras foram examinadas utilizando os métodos de microscopia óptica (MO), ensaio imunoenzimático (EI) e reação em cadeia da polimerase (PCR). As espécies e genótipos de Cryptosporidium foram determinados pelo método de polimorfismo no tamanho dos fragmentos de restrição (RFLP) ou sequenciamento dos genes SSU-rRNA e GP60. A infecção por Cryptosporidium spp. teve ocorrência de 14% (28/200). A ocorrência em bezerros (26%) foi significativamente maior do que em vacas (2%). Do total de 27 amostras positivas submetidas à caracterização genética, C. andersoni foi identificado em 23 (85.1%), C. bovis em três (11.1%) e C. parvum subtipo IIaA15G2R1 em uma (3.7%). O presente estudo demonstrou que a infecção por Cyptosporidium é comum e difundida em bovinos leiteiros nessa região e que bezerros possuem uma alta prevalência de C. andersoni. A presença de C. parvum subtipo IIaA15G2R1 indica que bezerros leiteiros dessa região devem ser considerados uma fonte de oocistos de Cryptosporidium com potencial zoonótico.

Cryptosporidium spp. and Giardia spp. in feces and water and the associated exposure factors on dairy farms

“…bovis , in line with other reports [36,61,64,65,66], but many researchers have detected these species more often when compared to C . parvum among cattle older than 2 months [44,53,59,66]. In others studies was suggested that infection with C .…”

“…This species is mainly identified among heifers and adult animals [44,63,69], but some investigators in Brazil identified C . andersoni at a high frequency in young calves too [66,70]. In our study, the prevalence of C .…”

Enteric Pathogens and Coinfections in Foals with and without Diarrhea

“…Nested PCR amplification was performed using the set of primers previously described for detection of Cryptosporidium spp. [36,37], G. duodenalis [38,39], and L. intracellularis DNA [8,40], with some modifications. Nested PCR products from the genera Cryptosporidium and Giardia with the expected sizes were analyzed by automated direct sequencing using a 3500 Genetic Analyzer (Applied Biosystems, Foster City, CA) and BigDye6 Terminator v3.1 Cycle Sequencing Kit.…”

Prevalence and characterization of Cryptosporidium spp. in dairy cattle in Nile River delta provinces, Egypt

“…106 Thus far, relative to the large number of studies in industrial-107 ized nations, only a few have been conducted to examine the dis-108 tribution of Cryptosporidium species in dairy cattle in developing 109 countries, where animals are managed very differently (Amer 110 et al, 2010;Inpankaew et al, 2010;Khan et al, 2010;Imre et al, 111 2011;Meireles et al, 2011;Muhid et al, 2011;wang et al, 112 2011a,b;Kang'ethe et al, 2012;Venu et al, 2012;Silva et al, 113 2013;Zhang et al, 2013). Most of these studies have shown a 114 common occurrence of C. bovis, C. ryanae, and C. andersoni in 115 pre-weaned calves, in addition to C. parvum Q3 (Meireles et al, 116 2011;Muhid et al, 2011;Wang et al, 2011a,b;Venu et al, 2012; 117 Silva et al, 2013;Zhang et al, 2013). A small number of these stud- approximately 830-bp fragment of the small subunit (SSU) rRNA 168 gene as previously described (Feng et al, 2007).…”