Prevalence of Fasciola infections and molecular identification of Fasciola in livestock in Dali Bai Autonomous Prefecture, Yunnan Province

doi:10.12140/j.issn.1000-7423.2024.06.004

Abstract

Abstract:

Objective To investigate the prevalence of Fasciola infections and to characterize the Fasciola species in livestock in Dali Bai Autonomous Prefecture. Methods Cattle, sheep, horse and pig stool samples were collected from 12 counties (city) in Dali Bai Autonomous Prefecture during the period from August 2023 to March 2024 for detection of Fasciola eggs. Parasite eggs were sampled from three Fasciola-infected stool specimens in each county, and photographed and measured with a microscopic imaging system. Then, genomic DNA was extracted from Fasciola eggs, and the ribosomal internal transcribed spacer (ITS) gene was amplified using a PCR assay for characterization of Fasciola species. The sequence of the ITS gene was aligned using the BLAST program in NCBI, and a neighbor-joining phylogenetic tree of Fasciola spp. was built based on the ITS gene sequence using the software MEGA 11.0. All statistical analyses were performed using the software SPSS 19.0, and differences in the prevalence of Fasciola infections were compared with chi-square test. Results A total of 1 302 livestock stool samples were collected, including 1 228 cattle stool samples (94.3%), 45 sheep stool samples (3.5%), 18 horse stool samples (1.4%), and 11 pig stool samples (0.8%), and there were 212 stool samples detected with suspected Fasciola eggs (16.3%). The detection of Fasciola eggs was 17.2% in cattle stool samples (211/1 228) and 2.2% in sheep stool samples (1/45), and no Fasciola eggs were detected in horse or pig stool samples. The highest detection of Fasciola eggs was seen in livestock stool samples from Weishan County (45.0%, 49/109), followed by from Eryuan County (42.7%, 44/103), Yunlong County (28.2%, 29/103), Midu County (25.4%, 30/118), Binchuan County (18.0%, 18/100), Heqing County (16.5%, 17/103), Xiangyun County (8.7%, 9/103), Jianchuan County (6.9%, 8/116), Nanjian County (4.7%, 5/107), Yangbi County (2.2%, 3/137), and no Fasciola eggs were detected in livestock stool samples from Dali City or Yongping County. There was a region-specific prevalence of Fasciola infections in livestock in Dali Bai Autonomous Prefecture (χ² = 222.975, P < 0.05), and the prevalence of Fasciola infections was higher in female livestock (19.0%, 165/873) than in males (11.0%, 47/429) (χ² = 13.319, P < 0.05). The eggs of F. gigantica and F. hepatica measured (182.30 ± 4.35) μm × (97.80 ± 5.11) μm and (149.49 ± 5.90) μm × (80.46 ± 4.37) μm, respectively. A total of 7 ITS gene sequences were obtained, including 2 from F. hepatica and 5 from F. gigantica, and these 7 sequences had been uploaded to the National Microbiology Data Center (NMDC), with NMDC numbers of NMDCN0005QTQ (sequence number: Binchuan County A1), NMDCN0005TN (sequence number: Weishan County A32), NMDCN0005QTO (sequence number: Jianchuan County A70), NMDCN0005ELF (sequence number: Yangbi County A19), NMDCN0005QTR (sequence number: Heqing County A7), NMDCN0005ELH (sequence number: Midu County A35) and NMDCN0005QTP (sequence number: Xiangyun County A18). Four sequences numbered Binchuan County A1, Weishan County A32, Jianchuan County A70 and Heqing County A7 had 100% sequence homology to those from F. gigantica (GenBank accession numbers: JF496711, OQ064778, MN608171 and MN97009), and the sequence numbered Xiangyun County A18 had 93% sequence homology to that from F. gigantica (GenBank accession number: MN97009), while two sequences numbered Yangbi County A19 and Weishan County A35 had 100% sequence homology to those from F. hepatica (GenBank accession number: JF432071 and JF496716). Phylogenetic analysis showed that the sequence numbered Yangbi A19 and sequences of F. hepatica from Australia (GenBank accession number: MN970007) and Iran (GenBank accession number: JF432071) were clustered into the same clade, and the sequence numbered Midu A35 and that of F. hepatica from Dali, China (GenBank accession number: JF496716) was clustered into the same clade, indicating a close genetic relationship. In addition, the remaining five sequences and those of F. gigantica from Dehong, China (GenBank accession number: JF496711), Bangladesh (GenBank accession number: OQ064778), Nigeria (GenBank accession number: MN608171), and Vietnam (GenBank accession numbers: MN970009 and MN970008) were clustered into the same clade, indicating a close genetic relationship. Conclusions The prevalence of Fasciola infections is high in livestock Dali Bai Autonomous Prefecture, Yunnan Province, and F. gigantica is the predominant species of Fasciola.

Key words: Fasciola spp., Molecular identification, Livestock, Dali

CLC Number:

R383.26

LI Shiyun, LI Cihuai, CHEN Feng, HAO Mingming, LI Xindi, ZHAO Yinjiao, ZHANG Dongqi, ZHANG Quanfu, LI Qincui, ZHANG Jianping, CHEN Shaorong, LUO Tianpeng, LIU Yuhua. Prevalence of Fasciola infections and molecular identification of Fasciola in livestock in Dali Bai Autonomous Prefecture, Yunnan Province[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2024, 42(6): 715-720.

Figures/Tables 3

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