Panmictic structure of the Cryptosporidium parvum population in Irish calves: influence of prevalence and host movement.

In total, 245 Cryptosporidium parvum specimens obtained from calves in 205 Irish herds between 2003 and 2005 were subtyped by sequencing the glycoprotein gene gp60 and performing multilocus analysis of seven markers. The transmission dynamics of C. parvum and the influence of temporal, spatial, parasitic, and host-related factors on the parasite (sub)populations were studied. The relationship of those factors to the risk of cryptosporidiosis was also investigated using results from 1,368 fecal specimens submitted to the veterinary laboratories for routine diagnosis during 2005. The prevalence was greatest in the northwest and midwest of the country and on farms that bought in calves. The panmixia (random mating) detected in the C. parvum population may relate to its high prevalence, the cattle density, and the frequent movement of cattle. However, local variations in these factors were reflected in the C. parvum subpopulations. This study demonstrated the importance of biosecurity in the control of bovine cryptosporidiosis (e.g., isolation and testing of calves before introduction into a herd). Furthermore, the zoonotic risk of C. parvum was confirmed, as most specimens possessed GP60 and MS1 subtypes previously described in humans.

Age-stratified Bayesian analysis to estimate sensitivity and specificity of four diagnostic tests for detection of Cryptosporidium oocysts in neonatal calves.

There is no gold standard diagnostic test for the detection of bovine cryptosporidiosis. Infection is usually highest in 2-week-old calves, and these calves also excrete high numbers of oocysts. These factors may give rise to variations in the sensitivity and specificity of the various diagnostic tests used to detect infection in calves of various ages. An age-stratified Bayesian analysis was carried out to determine the optimum diagnostic test to identify asymptomatic and clinical Cryptosporidium sp. infection in neonatal calves. Fecal samples collected from 82 calves at 1 week, 2 weeks, 3 weeks, and 4 weeks of age were subjected to the following tests: microscopic examination of smears stained with either phenol-auramine O or fluorescein isothiocyanate (FITC)-conjugated anti-Cryptosporidium monoclonal antibody, nested-PCR, and quantitative real-time PCR. The results confirmed a high prevalence of Cryptosporidium sp. infection, as well as a high level of oocyst excretion, in 2-week-old calves. The sensitivities of all the tests varied with the age of the calves. Quantitative real-time PCR proved to be the most sensitive and specific test for detecting infection irrespective of the age of the calf. The microscopic techniques were the least sensitive and exhibited only moderate efficiency with 2-week-old calves excreting large numbers of oocysts, the majority of which were diarrheic. It was concluded that, when interpreting the results of routine tests for bovine cryptosporidiosis, cognizance should be taken of the sensitivity of the tests in relation to the age of the calves and stage of infection.

Peri-parturient rise of Cryptosporidium oocysts in cows: new insights provided by duplex quantitative real-time PCR.

In order to clarify if a peri-parturient rise of Cryptosporidium parvum oocysts occurs in cows, faecal samples from 42 cows on two farms were collected. These samples were taken during the pre-parturient, the peri-parturient and the post-parturient periods. Two methods were used to detect the oocysts, a nested-PCR coupled with sequencing and a duplex real-time PCR (qPCR) that quantified Cryptosporidium spp. DNA concentration. The qPCR results were adjusted using a hierarchical Bayesian model taking into account within and between run variation. Generalised Estimating Equation models (GEE) were used to determine if peri-parturient cows were at greater risk of being infected than pre- or post-parturient cows. Fourteen dairy cows exhibited a peri-parturient and post-parturient rise in the excretion of Cryptosporidium spp. oocysts, other than the zoonotic C. parvum. The cows in the suckler beef farm were the only ones infected with the zoonotic species C. parvum at calving. Due to the low concentration of oocysts excreted mainly from species other than C. parvum, it would appear unlikely that cows act as a source of infection for their calves or contribute significantly to environmental contamination.