Microsatellite Typing of
            <i>Cryptosporidium parvum</i>
            in Isolates from a Waterborne Outbreak

Hunter, Paul R.; Wilkinson, Dawn C.; Lake, Iain R.; Harrison, Florence C.D.; Syed, Qutubuddin; Hadfield, Stephen J.; Chalmers, Rachel M.

doi:10.1128/jcm.01636-08

Cited by 16 publications

(10 citation statements)

References 4 publications

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“…The greatest number of alleles was detected at the ML2 locus (6 alleles), which provided a HGDI value even higher than gp60 sequencing. This microsatellite has been reported to be among the most polymorphic markers for typing C. parvum from humans, pre-weaned livestock or waterborne outbreaks in Europe (Cacciò et al, 2001;Hunter et al, 2007Hunter et al, , 2008Quílez et al, 2011Quílez et al, , 2013Díaz et al, 2012). The most prevalent allele at this marker (ML2-231) was also the most common allelic variant in calves in Spain (Quílez et al, 2011;Díaz et al, 2012;Ramo et al, 2016a).…”

Section: Discussionmentioning

confidence: 98%

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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Section: Discussionmentioning

confidence: 98%

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

et al. 2019

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“…Even where species identification has been undertaken, further analysis of isolates from cases may be required to investigate outbreaks. Retrospective analysis of C. parvum isolates by multilocus fragment typing enabled more accurate mapping of outbreak-related cases to water supply zones [68]. Some nine years after the Milwaukee outbreak, C. hominis isolates, further analysed for variation in the gp60 gene, were identified as indistinguishable from the outbreak isolate, year round in Milwaukee wastewater indicating continuing, stable transmission of human cryptosporidiosis in the city [69].…”

Section: Application Of Molecular Assays For Species Determination Anmentioning

confidence: 99%

Waterborne outbreaks of cryptosporidiosis

Chalmers

2012

Annali Dell'Istituto Superiore Di Sanità

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Abstract. Water is the most commonly reported vehicle of transmission in Cryptosporidium outbreaks. While mains drinking water quality is highly regulated in industrialised countries, treated recreational water venues remain highly variable and these have emerged as important settings in the transmission of cryptosporidiosis. Epidemiological investigations of outbreaks benefit from supplementary microbiological evidence and, more recently, the application of molecular typing data to link isolates from cases to each other and to suspected sources. This article documents how waterborne Cryptosporidium outbreaks are identified and reported, how such outbreaks have acted as drivers of regulatory change, and some of the recent developments in the detection and investigation of these outbreaks and their spread, especially the application of molecular typing assays.Key words: Cryptosporidium, genotyping, recreational, drinking, water.Riassunto (Epidemie di cryptosporidiosi trasmesse con le acque). L'acqua è il più comune veicolo di trasmissione di epidemie dovute a Cryptosporidium. Mentre la qualità delle acque potabili distribuite da acquedotti è fortemente regolamentata nei paesi industrializzati, la qualità delle acque trattate di ambienti ricreativi chiusi è fortemente variabile e questi ambienti si sono rivelati importanti nella trasmissione della criptosporidiosi. Le indagini epidemiologiche sulle epidemie traggono beneficio dalle evidenze microbiologiche e, più recentemente, dall'applicazione dei dati di tipizzazione molecolare per collegare i ceppi isolati con i casi d'infezione e con le fonti di contagio sospette. Quest'articolo documenta come vengono individuate e notificate le epidemie di Cryptosporidium trasmesse con le acque, come tali epidemie hanno agito da guida per migliorare la normativa, ed alcuni recenti sviluppi nella rilevazione e nelle indagini di queste epidemie e della loro diffusione, in particolare l'applicazione dei saggi di tipizzazione molecolare.Parole chiave: Cryptosporidium, genotipizzazione, acqua ad uso ricreativo, acqua potabile.

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“…Intraspecies investigations of C. parvum and C. hominis using multilocus typing approaches are helpful for further epidemiological and transmission studies (Mallon et al. 2003b; Hunter et al. 2007, 2008).…”

Section: Introductionmentioning

confidence: 99%

A whole water catchment approach to investigating the origin and distribution of Cryptosporidium species

Robinson

Chalmers

Stapleton

et al. 2011

Journal of Applied Microbiology

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Aims: Investigating the distribution and origin of Cryptosporidium species in a water catchment affected by destocking and restocking of livestock as a result of a foot and mouth disease epidemic. Methods and Results: Surface water, livestock and wildlife samples were screened for Cryptosporidium and oocysts characterised by sequencing SSU rRNA and COWP loci, and fragment analysis of ML1, ML2 and GP60 microsatellite loci. Oocyst concentrations in water samples (0–20·29 per 10 l) were related to rainfall events, amount of rainfall and topography. There was no detectable impact from catchment restocking. Cryptosporidium spp. found in water were indicative of livestock (Cryptosporidium andersoni and Cryptosporidium parvum) and wildlife (novel genotypes) sources. However, C. andersoni was not found in any animals sampled. Calf infections were age related; C. parvum was significantly more common in younger animals (<4 weeks old). Older calves shared Cryptosporidium bovis, Cryptosporidium ryanae and C. parvum. Wildlife shed C. parvum, Cryptosporidium ubiquitum, muskrat genotype II and deer genotype. Conclusions: Several factors affect the occurrence of Cryptosporidium within a catchment. In addition to farmed and wild animal hosts, topography and rainfall patterns are particularly important. Significance and Impact of the Study: These factors must be considered when undertaking risk‐based water safety plans.

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Microsatellite Typing of Cryptosporidium parvum in Isolates from a Waterborne Outbreak

Cited by 16 publications

References 4 publications

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

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

Waterborne outbreaks of cryptosporidiosis

A whole water catchment approach to investigating the origin and distribution of Cryptosporidium species

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