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[Isolation and identification of cow-origin Cryptosporidium isolates in Hefei]
Objective:
To isolate cow-origin Cryptosporidium in Hefei, and identify its species.
Methods:
285 dairy cattle fecal samples collected from a farm in Hefei were examined by using floating saturated solution of sucrose and modified acid-fast staining. Cryptosporidium oocysts were isolated and purified from positive fecal samples. Genetic DNA was extracted to be the template. According to the sequence of 18S rRNA gene and HSP70 gene from Cryptosporidium sp., the primers were designed and synthesized. The PCR products were amplified by PCR and nested-PCR. The nested PCR products were cloned and sequenced. Homology searches and phylogenic tree construction were done by DNAStar software.
Results:
Five fecal samples were positive by morphological methods with an infection rate of 1.8% (5/285). Oocysts from the 5 positive fecal samples were elliptical or ovoid detected by using floating saturated solution of sucrose and modified acid-fast staining with the size of 7.37 microm x 6.13 microm and 7.58 microm x 6.20 microm, and a shape index of 1.20 and 1.22, respectively. Nested-PCR resulted in a 18S rRNA and HSP70 gene fragments with approximately 250 bp and 325 bp, respectively. The five isolates showed a high level of nucleic acid identity with sequence data of the 18S rRNA gene of Cryptosporidium andersoni (DQ989573), and they were clustered in the same clade. The highest HSP70 gene sequence identity was found among the five isolates and other reported C. andersoni isolates (AY954892 and DQ989576), and they were placed into the same clade.
Conclusion:
The cow-origin Cryptosporidium isolates derived from Hefei is Cryptosporidium andersoni.
Insights
Cryptosporidium andersoni was identified in dairy cattle from Hefei, China. This study isolated and genetically identified cow-origin Cryptosporidium, confirming the presence of this parasite in the region.
Area of Science:
- Veterinary Parasitology
- Molecular Biology
- Microbiology
Context:
- Cryptosporidium is a significant enteric pathogen in cattle.
- Understanding the species prevalent in livestock is crucial for disease control.
- Previous studies on Cryptosporidium in Hefei cattle were limited.
Purpose:
- To isolate and identify the species of Cryptosporidium originating from cows in Hefei.
- To characterize the genetic makeup of these isolates.
Summary:
- Morphological and molecular methods (18S rRNA and HSP70 genes) were used to analyze 285 dairy cattle fecal samples.
- An infection rate of 1.8% was observed.
- Genetic analysis confirmed the isolates as Cryptosporidium andersoni, with high sequence identity to known strains.
Impact:
- This research identifies Cryptosporidium andersoni as the causative agent in Hefei cattle.
- Findings contribute to regional parasite surveillance and inform targeted intervention strategies.
- Establishes a baseline for future epidemiological studies on bovine cryptosporidiosis.
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