Experimental infection with Cryptosporidium parvum IIaA21G1R1 subtype in immunosuppressed mice

Coco, Valeria Fernanda Del; Córdoba, María Alejandra; Sidoti, Alicia; Santı́n, Mónica; Drut, Ricardo; Basualdo, Juan Ángel

doi:10.1016/j.vetpar.2012.06.033

Cited by 12 publications

(10 citation statements)

References 28 publications

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“…Infected mice began to shed small numbers of oocysts on 7-8 dpi, with two peaks in infection intensity during the exposure period. The oocysts lasted 27 days, consistent with previous studies [16,28,37], but different from the study of C. muris CB03 isolated from BALB/c mice infected with Camelus bactrianus [38]. The difference in the prepatent and patent periods may depend on the specific Cryptosporidium species and experimental animals [20].…”

Section: Discussionsupporting

confidence: 68%

Molecular identification and biological characterization of Cryptosporidium muris from camels (Camelus bactrianus) in China

et al. 2021

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Background Cryptosporidium is an opportunistic pathogen that infects a wide variety of vertebrates. The aim of the present study was to characterize Cryptosporidium spp. isolates from Bactrian camels and to foster further understanding of the biological characteristics of the pathogen. Methods Fecal specimens were collected from two 4-year-old Bactrian camels resident at the Kaifeng City Zoo in China and examined for Cryptosporidium. Fecal specimens were screened using the floatation method, and then genomic DNA was extracted from the oocysts and identified by nested-PCR amplification of the small subunit ribosomal RNA (SSU rRNA) gene, the actin gene and the Cryptosporidium oocyst wall-protein (COWP) gene. Subtype analysis was performed based on four minisatellite (MS) loci (MS1, MS2, MS3 and MS16) that were aligned and phylogenetically analyzed to determine the species and subtype of Cryptosporidium. We then established a BALB/c mice infection model and further verified the results through clinical status, pattern of oocyst excretion and histological examination. Results Cryptosporidium oocyst isolates from the two Bactrian camels had an average (± standard deviation) size of 7.49 ± 0.13 × 5.70 ± 0.10 μm (n = 50). The sequencing and phylogenetic analysis confirmed the species as C. muris. Multilocus sequence typing analysis indicated that the subtypes were M13, M4, M1 and M5. Following the inoculation of BALB/c mice, we found that the prepatent period and number of oocysts per gram increased with increasing infective dose. Oocysts were first detected in the feces of BALB/c mice at 7–8 days post-infection (dpi), with levels peaking twice thereafter, at 15–16 dpi and 19–20 dpi. Histology and scanning electron microscopy studies showed that the stomach contained gastric pits filled with Cryptosporidium that adhered to the surface of gastric mucosa gland epithelial cells, causing the latter to deform, swell and become disordered. Conclusions The findings of this study indicated that oocysts isolated from Bactrian camels were from C. muris. This is the first report of C. muris isolated from camels in China. More epidemiological data are needed to understand the prevalence and transmission of C. muris in camels in different geographic areas. Graphical abstract

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Section: Discussionsupporting

confidence: 68%

Molecular identification and biological characterization of Cryptosporidium muris from camels (Camelus bactrianus) in China

et al. 2021

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“…The subtype IIaA21G1R1 (n = 15) predominated in the study and was found to be frequent in the provinces of Buenos Aires (9 of 26), Cordoba (3 of 7), and Santa Fe (3 of 13). As to our knowledge, IIaA21G1R1 has so far only been identified in two calves from Sweden (Silverlås et al, 2010), and in a single calf from Argentina (Del Coco et al, 2012), and has not been reported in other regions of the world. The novel subtype IIaA23G1R1 (n = 7) was frequent in our study group predominating in Santa Fe (6 of 13) while it was identified only once in Buenos Aires (1 of 23).…”

Section: Resultsmentioning

confidence: 59%

Molecular characterization of Cryptosporidium isolates from calves in Argentina

Tomazic¹,

Maidana²,

Dominguez³

et al. 2013

Veterinary Parasitology

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“…They were experimentally infected using gastric tubes with a single dose of 1 × 10 5 Cryptosporidium oocysts 1 h before a meal. The oocysts were collected from naturally exposed buffalo calves [ 38 ]. After 4 days, we collected the fecal pellets of the infected mice every day for 3 weeks and examined them using MZN staining [ 33 ].…”

Section: Methodsmentioning

confidence: 99%

“…The first group comprised non-vaccinated non-infected mice (negative control). The mice in the second group were orally infected using gastric tubes with 1 × 10 5 C. parvum oocysts in 250 μL PBS (pH = 7.2) [ 38 ] 1 h before a meal on week 4 of the experiment (positive control). The mice in the third group were immunized with the Cryptosporidium -specific purified fraction by two doses of subcutaneous injection at 2-week intervals.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of a vaccine candidate isolated from Cryptosporidium parvum oocyst in mice

et al. 2022

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Background and Aim: Cryptosporidiosis is a leading cause of diarrheal disease worldwide and is an animal and public health burden. This study aimed to evaluate the protective potential of affinity-purified Cryptosporidium parvum oocyst antigen as a vaccine candidate according to fecal oocyst shedding, humoral and cellular immune responses, histopathological changes, and the number of parasite developmental stages in ileal and hepatic tissues. Materials and Methods: We isolated oocysts from naturally infected buffalo calves and identified them molecularly as C. parvum isolates (GenBank: ON730707 and ON730708) by targeting the Cryptosporidium oocyst wall protein gene. We propagated the C. parvum oocysts in mice. In addition, we prepared crude antigen from the isolated oocysts by purification using cyanogen bromide-activated Sepharose-4B affinity chromatography coupled with rabbit hyperimmune serum. Then, we divided 81 parasite-free mice into three groups: (1) non-vaccinated non-infected mice, (2) mice orally infected with 1 × 105 C. parvum oocysts on week 4 of the experiment, and (3) mice immunized twice with 40 μg/kg of the purified fraction at 2-week intervals. Then, we challenged the vaccinated group with C. parvum oocysts after 2 weeks, and the positive control group was infected at the same time. Results: We observed a prolonged prepatent period and decreased oocyst shedding in the vaccinated infected mice compared with the non-vaccinated infected mice (t < 0.001). The vaccinated mice had significantly higher immunoglobulin G levels than those in the other two groups at all examined weeks. In addition, the production of cytokines interferon-gamma, interleukin (IL)-10, IL-12, and IL-15 was activated post-vaccination. After the challenge, all tested cytokines were significantly increased (p < 0.001) in the two infected groups compared with the non-vaccinated non-infected group, with the highest levels in the vaccinated infected group. Vaccinated infected mice exhibited significantly fewer pathological lesions in the ileum and liver than non-vaccinated infected mice, which showed prominent histopathological lesions. Endogenous developmental stages of C. parvum indicated that the ileum was more parasitized than the liver and that vaccination resulted in a lower number of oocysts in ileal and hepatic tissues (p < 0.05). Conclusion: Our prepared affinity-purified vaccine candidate could be promising in protecting against cryptosporidiosis.

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Experimental infection with Cryptosporidium parvum IIaA21G1R1 subtype in immunosuppressed mice

Cited by 12 publications

References 28 publications

Molecular identification and biological characterization of Cryptosporidium muris from camels (Camelus bactrianus) in China

Molecular identification and biological characterization of Cryptosporidium muris from camels (Camelus bactrianus) in China

Molecular characterization of Cryptosporidium isolates from calves in Argentina

Evaluation of a vaccine candidate isolated from Cryptosporidium parvum oocyst in mice

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