Development of a PCR assay for detection and identification of Eimeria spp. in cattle

Eimeria spp. are important coccidian parasites causing diarrhea and significant mortality in cattle worldwide. To date, at least 13 Eimeria species with varying pathogenicity have been identified in cattle. Efficient detection and identification of Eimeria spp. is therefore essential for the prevent...

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Veröffentlicht in:Veterinary parasitology 2024-12, Vol.332, p.110315, Article 110315
Hauptverfasser: Chen, Xuehua, Deng, Miner, Chen, Nan, Chen, Xiaohong, Li, Na, Feng, Yaoyu, Xiao, Lihua, Guo, Yaqiong
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container_issue
container_start_page 110315
container_title Veterinary parasitology
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creator Chen, Xuehua
Deng, Miner
Chen, Nan
Chen, Xiaohong
Li, Na
Feng, Yaoyu
Xiao, Lihua
Guo, Yaqiong
description Eimeria spp. are important coccidian parasites causing diarrhea and significant mortality in cattle worldwide. To date, at least 13 Eimeria species with varying pathogenicity have been identified in cattle. Efficient detection and identification of Eimeria spp. is therefore essential for the prevention and control of bovine coccidiosis. However, the commonly used microscopic examination for Eimeria spp. is time-consuming and requires considerable expertise. In this study, we aligned the nucleotide sequences of the small subunit (SSU) rRNA gene of common Eimeria species and developed a nested PCR assay targeting the polymorphic SSU rRNA region of Eimeria spp. from cattle. Initially, the SSU rRNA gene PCR assay was compared with microscopic examination for sensitivity and detection range of Eimeria species using fecal samples from dairy cattle. Of the 193 fecal samples, 131 (67.9 %) and 78 (40.4 %) were positive for Eimeria by PCR and microscopy, respectively. Sequence analysis of the PCR products identified six Eimeria species, including E. cylindrica (n = 76), E. bovis (n = 54), E. auburnensis (n = 30), E. zuernii (n = 25), E. wyomingensis (n = 10), E. canadensis (n = 1), and co-infections of 2–4 species (n = 55). In contrast, only the first four species and co-infections of 2–3 species were identified by microscopy. The PCR assay was able to detect as few as 50 Eimeria oocysts per gram of feces. Thus, the developed SSU rRNA gene PCR assay has a high sensitivity and allowed easy identification of at least six common Eimeria species and their co-infections in cattle. It should be useful in molecular epidemiological studies of bovine coccidiosis. •A nested PCR assay was developed for detection and identification of Eimeria spp. in cattle.•The sensitivity threshold of the PCR assay was 50 Eimeria oocysts per gram of feces.•The PCR assay allowed identification of bovine Eimeria spp. and their co-infection.
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To date, at least 13 Eimeria species with varying pathogenicity have been identified in cattle. Efficient detection and identification of Eimeria spp. is therefore essential for the prevention and control of bovine coccidiosis. However, the commonly used microscopic examination for Eimeria spp. is time-consuming and requires considerable expertise. In this study, we aligned the nucleotide sequences of the small subunit (SSU) rRNA gene of common Eimeria species and developed a nested PCR assay targeting the polymorphic SSU rRNA region of Eimeria spp. from cattle. Initially, the SSU rRNA gene PCR assay was compared with microscopic examination for sensitivity and detection range of Eimeria species using fecal samples from dairy cattle. Of the 193 fecal samples, 131 (67.9 %) and 78 (40.4 %) were positive for Eimeria by PCR and microscopy, respectively. Sequence analysis of the PCR products identified six Eimeria species, including E. cylindrica (n = 76), E. bovis (n = 54), E. auburnensis (n = 30), E. zuernii (n = 25), E. wyomingensis (n = 10), E. canadensis (n = 1), and co-infections of 2–4 species (n = 55). In contrast, only the first four species and co-infections of 2–3 species were identified by microscopy. The PCR assay was able to detect as few as 50 Eimeria oocysts per gram of feces. Thus, the developed SSU rRNA gene PCR assay has a high sensitivity and allowed easy identification of at least six common Eimeria species and their co-infections in cattle. It should be useful in molecular epidemiological studies of bovine coccidiosis. •A nested PCR assay was developed for detection and identification of Eimeria spp. in cattle.•The sensitivity threshold of the PCR assay was 50 Eimeria oocysts per gram of feces.•The PCR assay allowed identification of bovine Eimeria spp. and their co-infection.</description><identifier>ISSN: 0304-4017</identifier><identifier>ISSN: 1873-2550</identifier><identifier>EISSN: 1873-2550</identifier><identifier>DOI: 10.1016/j.vetpar.2024.110315</identifier><identifier>PMID: 39270603</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Animals ; Cattle ; Cattle Diseases - diagnosis ; Cattle Diseases - epidemiology ; Cattle Diseases - parasitology ; Coccidiosis - diagnosis ; Coccidiosis - epidemiology ; Coccidiosis - parasitology ; Coccidiosis - veterinary ; Detection ; DNA, Protozoan - genetics ; Eimeria - classification ; Eimeria - genetics ; Eimeria - isolation &amp; purification ; Eimeria spp ; Feces - parasitology ; Identification ; Polymerase Chain Reaction - methods ; Polymerase Chain Reaction - veterinary ; Sensitivity and Specificity ; SSU rRNA gene PCR</subject><ispartof>Veterinary parasitology, 2024-12, Vol.332, p.110315, Article 110315</ispartof><rights>2024</rights><rights>Copyright © 2024. 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Sequence analysis of the PCR products identified six Eimeria species, including E. cylindrica (n = 76), E. bovis (n = 54), E. auburnensis (n = 30), E. zuernii (n = 25), E. wyomingensis (n = 10), E. canadensis (n = 1), and co-infections of 2–4 species (n = 55). In contrast, only the first four species and co-infections of 2–3 species were identified by microscopy. The PCR assay was able to detect as few as 50 Eimeria oocysts per gram of feces. Thus, the developed SSU rRNA gene PCR assay has a high sensitivity and allowed easy identification of at least six common Eimeria species and their co-infections in cattle. 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subjects Animals
Cattle
Cattle Diseases - diagnosis
Cattle Diseases - epidemiology
Cattle Diseases - parasitology
Coccidiosis - diagnosis
Coccidiosis - epidemiology
Coccidiosis - parasitology
Coccidiosis - veterinary
Detection
DNA, Protozoan - genetics
Eimeria - classification
Eimeria - genetics
Eimeria - isolation & purification
Eimeria spp
Feces - parasitology
Identification
Polymerase Chain Reaction - methods
Polymerase Chain Reaction - veterinary
Sensitivity and Specificity
SSU rRNA gene PCR
title Development of a PCR assay for detection and identification of Eimeria spp. in cattle
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