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

Abstract

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

Microbiome-Metabolomics Analysis of the Impacts of Cryptosporidium muris Infection in BALB/C Mice

“…Conversely, significantly increased ACE index ( P < 0.01) and Chao index ( P < 0.01) at 14 days after C. muris infection indicated that the fecal-microbiota richness was increased during this period. Overall, these data suggested that mice infected with C. muris had higher microbiota richness at the peak of oocysts shedding (14 dpi) ( 5 ). β-Diversity analysis was used to confirm the differences and similarities in gut microbial community composition between groups.…”

“…Bacteria such as Lachnospiraceae and Desulfovibrionaceae increased in abundance during C. muris infection, especially at 14 dpi. Interestingly, previous studies have shown that the number of shed oocysts peaks on the second week after C. muris infection in mice ( 5 , 21 ). Lachnospiraceae contribute toward to pyruvate metabolism to drive SCFA biosynthesis ( 22 ).…”

“…It is a zoonotic parasite discovered by Tyzzer in the stomach glands of laboratory mice in 1907 ( 2 , 3 ). The life cycle of C. muris is similar to other Cryptosporidium species, but the infection site of C. muris is the stomach rather than the intestinal mucosal epithelial cells ( 4 , 5 ). The diarrhea induced by Cryptosporidium is caused by significant mucosal inflammation, reduced absorption surface area, and malabsorption ( 6 ).…”

An Update on Zoonotic Cryptosporidium Species and Genotypes in Humans

“…Cryptosporidium muris , a gastric species, has a very wide host range including rodents, ruminants, cats and dogs, horses, Bactrian camels, pigs, birds, and non-human primates [ 22 , 23 , 40 , 162 , 194 , 195 , 196 , 197 , 198 ]. There have been numerous reports in humans, particularly in developing countries (in children and HIV patients) [ 1 , 5 ], ( Table 1 ).…”

Therapeutic Potency of Ginger, Garlic, and Pomegranate Extracts Against Cryptosporidium parvum-Mediated Gastro-Splenic Damage in Mice

“…Light microscopic examination of C. parvum-infected gastric sections showed desquamation of the gastric glands' epithelium associated with the presence of oocysts within the epithelial lining and within the lumen of the gastric glands as well as marked hyperplastic changes within the mucus cells [33]. Concerning the gastric sections of the infected MTZ, garlic, ginger, and pomegranate-treated mice, they retrieved their normal structure-like healthy noninfected mice.…”