Multilocus typing of Cryptosporidium spp. in young calves with diarrhea in Korea

- Cryptosporidium spp. were detected using PCR and ELISA in diarrheal feces from calves.
- PCR and ELISA data showed good agreement in detecting C. parvum.
- Multilocus typing with 18S rRNA, gp60, and hsp70 genes of C. parvum was performed.
- Subtyping of C. parvum gp60 gene first revealed IIaA18G3R1 and IIaA16G3R1 in Asia.
- PCR and sequencing of hsp70 gene clearly differentiated C. bovis and C. ryanae.

We assessed the prevalence and performed molecular analysis of Cryptosporidium spp. in diarrheal feces of calves in Korea. Diarrheal fecal samples were collected from 951 young calves (<3 months) on 425 farms. Cryptosporidium prevalence was assessed by PCR and ELISA, and molecular characterization was performed by targeting the 18S rRNA, heat-shock protein 70 (hsp70), and glycoprotein 60 (gp60) genes. Data were analyzed according to the sex, type of cattle, region, season, and type of diarrhea. PCR analysis revealed Cryptosporidium spp. in 9.9% (94/951) of diarrheal fecal samples. C. parvum and C. bovis/ryanae were present in 6.1% (58/951) and 4.1% (39/951) of diarrheal fecal samples, respectively. In addition, ELISA showed positive results for C. parvum in 9.7% (92/951) samples. Statistical analysis of the PCR and ELISA results revealed a lower prevalence of C. parvum in the hemorrhagic diarrheal samples (P < 0.05). For C. bovis/ryanae, seasonality and high prevalence in hemorrhagic diarrhea were observed (P < 0.05). Of the 951 samples tested for C. parvum, 903 samples showed agreement with a κ value of 0.65, indicating good agreement between the two tests. Although C. bovis and C. ryanae share highly similar 18S rRNA sequences, PCR based on hsp70 successfully distinguished C. bovis from C. ryanae. Sequence analysis of gp60 revealed that C. parvum belonged to the IIa families and was further subtyped as IIaA18G3R1 and IIaA16G3R1, which have not been previously reported in Asia. These findings indicate that Cryptosporidium spp. play an important role in diarrhea in young calves in Korea. Considering the zoonotic significance of C. parvum IIa subtype and dense rearing system of cattle in Korea, prevention and continuous monitoring of Cryptosporidium are required.