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Identification of Zoonotic Balantioides coli in Pigs by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and Its Distribution in Korea.

ABSTRACT: Balantioides coli is a zoonotic protozoan parasite whose main reservoir is pigs. Recent studies have shown that B. coli variant A but not B has zoonotic potential. While B. coli infection has been reported in different animals and countries, the prevalence of the zoonotic variant is limited due to a lack of molecular information. Therefore, this study investigated the prevalence of B. coli in domestic pigs in Korea and assessed its zoonotic potential. A total of 188 pig fecal samples were collected from slaughterhouses in Korea. B. coli was identified by microscopy and molecular methods. B. coli was identified in 79 (42.9%) and 174 (94.6%) samples by microscopy and polymerase chain reaction (PCR), respectively. This study also developed a PCR-restriction fragment length polymorphism (PCR-RFLP) method to differentiate B. coli variant A from B without sequence analysis. Using this method, 62 (33.7%) and 160 (87.0%) samples were positive for variants A and B, respectively, and 48 (26.1%) samples were co-infected with both variants. Sequence and phylogenetic analyses showed a high genetic diversity of B. coli in pigs in Korea. To our knowledge, this is the first study to develop a method to differentiate B. coli variants A and B without sequence analysis and to assess the molecular epidemiology of B. coli in pigs. Continuous monitoring of zoonotic B. coli in pigs should be performed as pigs are the main source of human balantidiasis.

SUBMITTER: Byun JW

PROVIDER: S-EPMC8465230 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:The zoonotic bacteria Capnocytophaga canimorsus and C. cynodegmi, the predominant Capnocytophaga species in the canine oral biota, can cause human local wound infections or lethal sepsis, usually transmitted through dog bites. Molecular surveying of these Capnocytophaga species using conventional 16S rRNA-based PCR is not always accurate due to their high genetic homogeneity. In this study, we isolated Capnocytophaga spp. from the canine oral cavity and identified them using 16S rRNA and phylogenetic analysis. A novel 16S rRNA PCR-restriction fragment length polymorphism (RFLP) method was designed based on our isolates and validated using published C. canimorsus and C. cynodegmi 16S rRNA sequences. The results showed that 51% of dogs carried Capnocytophaga spp. Among these, C. cynodegmi (47/98, 48%) was the predominant isolated species along with one strain of C. canimorsus (1/98, 1%). Alignment analysis of 16S rRNA sequences revealed specific site nucleotide diversity in 23% (11/47) of the C. cynodegmi isolates, which were misidentified as C. canimorsus using previously reported species-specific PCR. Four RFLP types could be classified from all the isolated Capnocytophaga strains. The proposed method demonstrates superior resolution in distinguishing C. cynodegmi (with site-specific polymorphism) from C. canimorsus and especially in distinguishing C. canimorsus from other Capnocytophaga species. After in silico validation, this method was revealed to have an overall detection accuracy of 84%; notably, accuracy reached 100% in C. canimorsus strains isolated from human patients. Overall, the proposed method is a useful molecular tool for the epidemiological study of Capnocytophaga in small animals and for the rapid diagnosis of human C. canimorsus infections. IMPORTANCE With the increased number of small animal breeding populations, zoonotic infections associated with small animals need to be taken more seriously. Capnocytophaga canimorsus and C. cynodegmi are part of common biota in the mouths of small animals and can cause human infections through bites or scratches. In this study, C. cynodegmi with site-specific 16S rRNA sequence polymorphisms was erroneously identified as C. canimorsus during the investigation of canine Capnocytophaga by conventional PCR. Consequently, the prevalence of C. canimorsus is incorrectly overestimated in epidemiological studies in small animals. We designed a new 16S rRNA PCR-RFLP method to accurately distinguish zoonotic C. canimorsus from C. cynodegmi. After validation against published Capnocytophaga strains, this novel molecular method had high accuracy and could detect 100% of C. canimorsus-strain infections in humans. This novel method can be used for epidemiological studies and the diagnosis of human Capnocytophaga infection following exposure to small animals.

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Identification of Zoonotic Balantioides coli in Pigs by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and Its Distribution in Korea.

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