Molecular characterisation and risk factor analysis of Cryptosporidium spp. in calves from Italy

Díaz, Pablo; Varcasia, Antonio; Pipia, Anna Paola; Tamponi, Claudia; Sanna, G.; Prieto, Alberto; Ruiu, Angelo; Spissu, P; Díez‐Baños, P.; Morrondo, P.; Scala, Antonio

doi:10.1007/s00436-018-6000-x

Cited by 57 publications

(65 citation statements)

References 58 publications

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“…All of the sequenced Cryptosporidium samples were confirmed as C. parvum subtype family IIa, which is in accordance with results from other countries as Canada [27], USA [28], England [29], Spain [25], New Zealand [30], Italy [31] and Germany [58]. Subtype family IId, common in Italian calves [31], was not detected in the present study. Specifically, 82.9% of the sequenced samples revealed C. parvum subtype IIaA15G2R1, the most prevalent subtype in calves worldwide [32,58].…”

Section: Discussionsupporting

confidence: 92%

Molecular characterization of Giardia intestinalis and Cryptosporidium parvum from calves with diarrhoea in Austria and evaluation of point-of-care tests

Lichtmannsperger

Hinney

Joachim

et al. 2019

Comparative Immunology, Microbiology and Infectious Diseases

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A B S T R A C TTo obtain information about the occurrence and genotype distribution of G. intestinalis and C. parvum in Austrian cattle, faecal samples from diarrhoeic calves younger than 180 days of age originating from 70 farms were examined. Of the 177 faecal samples, 27.1% were positive for Giardia cysts (immunofluorescence microscopy) and 55.4% for Cryptosporidium oocysts (phase-contrast microscopy). Positive samples were characterized by nested PCR for Giardia, 83.3% (triosephosphate isomerase; tpi) and 89.6% (β-giardin; bg) were positive, while the Cryptosporidium nested PCR returned 92.5% (60-kDa glycoprotein) positive results. Sequence analysis revealed one assemblage A-positive sample and 30 (bg) respectively 29 (tpi) assemblage E-positive samples for G. intestinalis. For C. parvum four subtypes within the IIa family (IIaA15G2R1, n = 29; IIaA19G2R2, n = 3; IIaA21G2R1, n = 2; IIaA14G1R1, n = 1) could be differentiated. Validation of two immunochromatographic point-of-care tests resulted in a sensitivity of 29.2% and 77.6%; a specificity of 98.4% and 91.1% for the detection of Giardia intestinalis and Cryptosporidium parvum, respectively. Results confirm the widespread occurrence of both protozoa in diarrhoeic calves in Austria.

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Section: Discussionsupporting

confidence: 92%

Molecular characterization of Giardia intestinalis and Cryptosporidium parvum from calves with diarrhoea in Austria and evaluation of point-of-care tests

Lichtmannsperger

Hinney

Joachim

et al. 2019

Comparative Immunology, Microbiology and Infectious Diseases

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“…In contrast to results from China (Cai et al, 2017;Feng and Xiao, 2017), the occurrence of this zoonotic IId family group in calves is rare in Europe and seems likely to have spread from Western Asia to other regions including France (Wang et al, 2014). This subtype family was described in European countries, such as in calves from Italy (Díaz et al, 2018), Belgium (Geurden et al, 2007), but never previously reported in France. Thus, the present study is the first to report the presence of the IId subtype in French calves.…”

Section: Discussioncontrasting

confidence: 68%

Molecular characterization of Cryptosporidium isolates from diarrheal dairy calves in France

Mammeri

Chevillot

Chenafi

et al. 2019

Veterinary Parasitology: Regional Studies and Reports

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A B S T R A C TCryptosporidium is an obligate intracellular protist parasite infecting a wide range of vertebrate hosts and causes significant intestinal disease in both animals and humans, as some species are zoonotic. Cattle and especially calves have been identified as one of the most common reservoirs of this protist. However, little is known about the genetics of Cryptosporidium in calves in some regions of France. The aim of this study was to detect and isolate Cryptosporidium spp. in faecal samples from naturally infected pre-weaned calves (≤45 days-old) in France. A total of 35 diarrhoeic pre-weaned calf faecal samples were collected from 26 dairy cattle farms in six departments (French administrative provinces). Cryptosporidium presence was established by microscopically screening samples for oocystes with an immunofluorescent (DFA) staining method. DFA-positive samples were then analysed by PCR-RFLP and 18S rRNA gene sequencing to determine species. Cryptosporidium parvum-positive samples were subtyped via nested PCR analysis of a partial fragment of the 60 kDa glycoprotein (gp60) gene product. Data were then integrated into phylogenetic tree analysis. DFA revealed the presence of Cryptosporidium oocysts in 31 out of 35 (88%) samples. Combined with 18S rRNA gene analysis results, C. parvum was detected in 30 samples. Subtyping analysis in 27/30 samples (90%) of the C. parvum isolates revealed two zoonotic subtype families, IIa (24/27) and IId (3/27). Four subtypes were recognised within the subtype family IIa, including the hypertransmissible IIaA15G2R1 subtype that is the most frequently reported worldwide (21/ 27), IIaA17G3R1 (1/27), IIaA17G1R1 (1/27), and IIaA19G1R1 (1/27). Two subtypes were recognised within the IId subtype family including IIdA22G1 (2/27) and IIdA27G1 (1/27). These findings illustrate the high occurrence of Cryptosporidium in calves in dairy herds and increase the diversity of molecularly characterised C. parvum isolates with the first description of IIaA17G3R1, IIaA19G1R1, and IId subtypes in France. The presence of zoonotic C. parvum subtype families (IIa, IId) in this study suggests that pre-weaned calves are likely to be a significant reservoir of zoonotic C. parvum, and highlights the importance of animal to human cryptosporidiosis transmission risk. Further molecular studies in calves and small ruminants from other French regions are required to better understand the epidemiology of cryptosporidiosis in France.However, due to strong inter-species similarities in microscopic size and shape, species within this genus require additional identification via genetic characterization. Molecular biology techniques have enabled the description of species that are highly host-specific, as well as others that are capable of infecting many hosts. Cryptosporidium parvum is considered to be the most prevalent species worldwide and a major zoonotic transmission risk (Xiao, 2010). Using molecular approaches to genetically characterise Cryptosporidium spp. has facilitated an

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“…However, most of these studies relied on microscopic examination of fecal specimens. Cryptosporidium infection rates in Europe have been reported as follows: 38.8% of calves in Italy [3], 37% and 12% of dairy and beef calves, respectively, in Belgium [37], 36.7% of calves in Sweden [38], 22.5% of calves in Poland [39], 19.2% of lambs and 37.1% of goats in Poland [12], 74.4% of lambs and 93.8% of goat kids in Spain [11], 13.1% of lambs and 9.5% of goat kids in Belgium [40], and 5.1% of lambs and 7.1% of goat kids in Greece [10]. From these studies, it is clear that cryptosporidial infections are widespread in calves, lambs, and goat kids in Europe, including Turkey.…”

Section: Discussionmentioning

confidence: 99%

“…Cryptosporidium parvum has a number of subtypes that can be grouped into subtype families as measured by gp60 sequence analyses [2]. Healthy and diarrheic calves less than one month of age may facilitate the transmission of cryptosporidiosis in both humans and animals [3]. Other livestock are also potential reservoirs of this protozoan.…”

Section: Introductionmentioning

confidence: 99%

Molecular detection of genotypes and subtypes of Cryptosporidium infection in diarrheic calves, lambs, and goat kids from Turkey

Kabir

Ceylan

et al. 2020

Parasitology International

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Background: Cryptosporidium spp. are enteric protozoan parasites that infect a wide range of animals as well as humans. The studies on Cryptosporidium infections of animals in Turkey are mostly rely on microscopic observation. Few data are available regarding the distribution of Cryptosporidium genotypes and subtypes infection. The aim of this study is to analyse the zoonotic potential of Cryptosporidium oocysts shed from young ruminant livestock. Methods: A total of 415 diarrheic fecal specimens from 333 calves, 67 lambs, and 15 goat kids were examined for the presence of Cryptosporidium oocysts by microscopy. Microscopic positive specimens were then analyzed for Cryptosporidium genotypes and subtypes detection by use of nested PCR of the small subunit ribosomal RNA (SSU rRNA) gene and the highly polymorphic 60 kDa glycoprotein (gp60) gene followed by sequence analyses. Results: The results of this study revealed that 25.6% (106 of 415) of the specimens were positive for Cryptosporidium spp. infection by microscopic examination and molecular analysis. We identified 27.4% (91/333), 19.4% (13/67), and 13.4% (2/15) of positivity in calves, lambs and goat kids, respectively. Genotyping of the SSU rRNA indicated that almost all positive specimens were of C. parvum , except for one calf which was of C. bovis. Sequence analysis of the gp60 gene revealed the most common zoonotic subtypes (IIa and IId) of C. parvum. We detected 11 subtypes (IIaA11G2R1,

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Molecular characterisation and risk factor analysis of Cryptosporidium spp. in calves from Italy

Cited by 57 publications

References 58 publications

Molecular characterization of Giardia intestinalis and Cryptosporidium parvum from calves with diarrhoea in Austria and evaluation of point-of-care tests

Molecular characterization of Giardia intestinalis and Cryptosporidium parvum from calves with diarrhoea in Austria and evaluation of point-of-care tests

Molecular characterization of Cryptosporidium isolates from diarrheal dairy calves in France

Molecular detection of genotypes and subtypes of Cryptosporidium infection in diarrheic calves, lambs, and goat kids from Turkey

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