A multiplex PCR test to identify four common cattle-adapted Cryptosporidium species

Thomson, Sarah; Innes, Elisabeth A.; Jonsson, Nicholas N.; Katzer, Frank

doi:10.1017/pao.2016.2

Cited by 17 publications

(25 citation statements)

References 24 publications

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“…This method therefore captured the severity of the disease by including only the disease timeframe (G. Innocent, BiOSS, Personal Communication). In addition, Bueno da Silva et al (2019) used microscopy to determine oocyst concentration, whereas the current study used 18S nested species-specific PCR (Thomson et al, 2016), allowing speciation of Cryptosporidium and determination that the oocysts present were C. parvum. The results from this study are in contrast to a study looking at growth rates in dairy calves with and without diarrhoea in New York (Virtala et al, 1996), which found no significant difference in growth rates between the two groups of calves.…”

Section: Discussionmentioning

confidence: 99%

“…Cryptosporidium spp. were identified using a multiplex PCR (Thomson et al, 2016), amplifying the 18S region, including specific primers for species commonly found in cattle, namely C. parvum, Cryptosporidium bovis, Cryptosporidium ryanae and Cryptosporidium andersoni. Any C. parvumpositive samples were genotyped at the gp60 gene and loci MM5, MM18, MM19 and TP14 (Morrison et al, 2008;Hotchkiss et al, 2015) to determine the multilocus genotypes of the affected calves.…”

Section: Diagnosis and Genotyping Of Cryptosporidiummentioning

confidence: 99%

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Long-term production effects of clinical cryptosporidiosis in neonatal calves

Shaw

Innes

Morrison

et al. 2020

International Journal for Parasitology

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Cryptosporidiosis can have a devastating effect in neonatal calves, resulting in diarrhoea, dehydration and, in severe cases, death of the animal. The disease is caused by Cryptosporidium spp. and is one of the most common causes of calf enteritis in the UK. The parasite is very difficult to remove from the farm, as the oocysts have a tough outer wall which enables the parasite to survive for several months in moist temperate environmental conditions and it is difficult to kill oocysts with common disinfectants used on a farm. If appropriate management practises are applied, the disease is usually self-limiting and most calves will recover. It has been shown, in studies with children and in lambs, that severe clinical cryptosporidiosis can result in long-term growth and cognitive impairment compared with individuals with no obvious signs of the disease. This study measured the long-term growth rate of beef calves on farm by comparing groups of animals that had suffered differing degrees of clinical severity of cryptosporidiosis as neonates. A group of 27 beef calves were enrolled in the study and monitored from birth to 6 months of age. The calves were scored for severity of cryptosporidiosis and weighed at regular intervals. The average difference in weight gain, at 6 months, between a group of calves that had severe cryptosporidiosis as neonates and a group of calves with no clinical signs of infection was 34 kg. Those calves that had experienced severe cryptosporidiosis as neonates showed a significantly reduced live weight gain compared with those calves showing no clinical signs of infection (P = 0.034). Therefore, the impact of severe cryptosporidiosis in neonatal calves has longer term effects on weight gain and production efficiency, resulting in the parasite having a greater impact on cattle production than previously thought.

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Section: Discussionmentioning

confidence: 99%

Section: Diagnosis and Genotyping Of Cryptosporidiummentioning

confidence: 99%

Long-term production effects of clinical cryptosporidiosis in neonatal calves

Shaw

Innes

Morrison

et al. 2020

International Journal for Parasitology

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“…Amplification of DNA was by nested PCR targeting the 18S gene [19]. Briefly, each 25 μL reaction contained 10 × PCR buffer (45 mM Tris–HCl pH 8.8, 11 mM (NH4)2SO 4 , 4.5 mM MgCl 2 , 4.4 μM EDTA, 113 μg mL −1 BSA, 1 mM each of four deoxyribonucleotide triphosphates), 0.5 units BioTaq (BIO-21040, Bioline, London, UK) and 10 μM of each primer.…”

Section: Methodsmentioning

confidence: 99%

Cryptosporidium Prevalence in Calves and Geese Co-Grazing on Four Livestock Farms Surrounding Two Reservoirs Supplying Public Water to Mainland Orkney, Scotland

et al. 2019

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The parasite Cryptosporidium parvum represents a threat to livestock health and production, water quality and public health. Cattle are known to be significant reservoirs of C. parvum, but transmission routes are complex and recent studies have implicated the potential role of wildlife in parasite transmission to cattle and water sources. On the Orkney Isles, high densities of Greylag geese (Anser anser) cause widespread faecal contamination of cattle pastures, where cryptosporidiosis is known to be the main cause of neonatal calf diarrhoea and Cryptosporidium contamination frequently occurs in two reservoirs supplying Mainland Orkney’s public water. This study aimed to determine the Cryptosporidium species and subtypes present in geese and calves co-grazing on four farms surrounding two reservoirs on Mainland Orkney. Results indicated a high level of C. parvum prevalence in calves, geese and water samples. gp60 analysis illustrated that higher genotypic diversity was present in the goose population compared with calves, but did not yield sequence results for any of the water samples. It can be concluded that the high levels of C. parvum evident in calves, geese and water samples tested represents a significant risk to water quality and public health.

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“…Molecular assays are essential to give further information on parasite species and genotype [38]. Different Cryptosporidium species can be detected using multiplex PCR assays [39] or by direct sequencing but this is not done as a routine diagnostic test. Molecular-based diagnostics are widely used in commercial labs, often using real time PCR (that is helpful to detect low concentrations of oocysts) and are increasingly used in human diagnostics.…”

Section: Transmission Of Cryptosporidiummentioning

confidence: 99%