Biochemical and molecular characterization of Campylobacter fetus isolates from bulls subjected to bovine genital campylobacteriosis diagnosis in Spain
Abstract Background Bovine genital campylobacteriosis (BGC) is caused by Campylobacter fetus subsp. venerealis (Cfv) including its biovar intermedius (Cfvi). This sexually transmitted disease induces early reproductive failure causing considerable economic losses in the cattle industry. Using a coll...
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| Format: | Article |
| Language: | English |
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BMC
2024-04-01
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| Series: | BMC Veterinary Research |
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| Online Access: | https://doi.org/10.1186/s12917-024-03970-8 |
| _version_ | 1827291528097169408 |
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| author | Nerea Pena-Fernández Nekane Kortabarria Ana Hurtado Medelin Ocejo Marcelo Fort Iratxe Pérez-Cobo Esther Collantes-Fernández Gorka Aduriz |
| author_facet | Nerea Pena-Fernández Nekane Kortabarria Ana Hurtado Medelin Ocejo Marcelo Fort Iratxe Pérez-Cobo Esther Collantes-Fernández Gorka Aduriz |
| author_sort | Nerea Pena-Fernández |
| collection | DOAJ |
| description | Abstract Background Bovine genital campylobacteriosis (BGC) is caused by Campylobacter fetus subsp. venerealis (Cfv) including its biovar intermedius (Cfvi). This sexually transmitted disease induces early reproductive failure causing considerable economic losses in the cattle industry. Using a collection of well-characterized isolates (n = 13), C. fetus field isolates (n = 64) and saprophytic isolates resembling Campylobacter (n = 75) obtained from smegma samples of breeding bulls, this study evaluated the concordance of the most used phenotypic (H2S production in cysteine medium and 1% glycine tolerance) and molecular (PCR) methods for the diagnosis of BGC and assessed possible cross-reactions in the molecular diagnostic methods. Results Characterization at the subspecies level (fetus vs. venerealis) of C. fetus isolated from bull preputial samples using phenotypic and molecular (PCR targeting nahE and ISCfe1) methods showed moderate concordance (κ = 0.462; CI: 0.256–0.669). No cross-reactions were observed with other saprophytic microaerophilic species or with other Campylobacter species that can be present in preputial samples. Whole genome sequencing (WGS) of discrepant isolates showed 100% agreement with PCR identification. For the differentiation of Cfv biovars, comparison of the H2S test (at 72 h and 5 days of incubation) and a PCR targeting the L-cysteine transporter genes showed higher concordance when H2S production was assessed after 5 days (72 h; κ = 0.553, 0.329–0.778 CI vs. 5 days; κ = 0.881, 0.631–1 CI), evidencing the efficacy of a longer incubation time. Conclusions This study confirmed the limitations of biochemical tests to correctly identify C. fetus subspecies and biovars. However, in the case of biovars, when extended incubation times for the H2S test (5 days) were used, phenotypic identification results were significantly improved, although PCR-based methods produced more accurate results. Perfect agreement of WGS with the PCR results and absence of cross-reactions with non-C. fetus saprophytic bacteria from the smegma demonstrated the usefulness of these methods. Nevertheless, the identification of new C. fetus subspecies-specific genes would help to improve BGC diagnosis. |
| first_indexed | 2024-04-24T12:38:34Z |
| format | Article |
| id | doaj.art-96fd7da8c996474c8a406b6705a86284 |
| institution | Directory Open Access Journal |
| issn | 1746-6148 |
| language | English |
| last_indexed | 2024-04-24T12:38:34Z |
| publishDate | 2024-04-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Veterinary Research |
| spelling | doaj.art-96fd7da8c996474c8a406b6705a862842024-04-07T11:21:46ZengBMCBMC Veterinary Research1746-61482024-04-0120111210.1186/s12917-024-03970-8Biochemical and molecular characterization of Campylobacter fetus isolates from bulls subjected to bovine genital campylobacteriosis diagnosis in SpainNerea Pena-Fernández0Nekane Kortabarria1Ana Hurtado2Medelin Ocejo3Marcelo Fort4Iratxe Pérez-Cobo5Esther Collantes-Fernández6Gorka Aduriz7SERIDA, Servicio Regional de Investigación y Desarrollo AgroalimentarioDepartment of Animal Health, NEIKER-Basque Institute for Agricultural Research and Development, Basque Research and Technology Alliance (BRTA), Parque Científico y Tecnológico de Bizkaia, P812Department of Animal Health, NEIKER-Basque Institute for Agricultural Research and Development, Basque Research and Technology Alliance (BRTA), Parque Científico y Tecnológico de Bizkaia, P812Department of Animal Health, NEIKER-Basque Institute for Agricultural Research and Development, Basque Research and Technology Alliance (BRTA), Parque Científico y Tecnológico de Bizkaia, P812SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of MadridCentral Veterinary-Animal Health Laboratory (LCV)SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of MadridDepartment of Animal Health, NEIKER-Basque Institute for Agricultural Research and Development, Basque Research and Technology Alliance (BRTA), Parque Científico y Tecnológico de Bizkaia, P812Abstract Background Bovine genital campylobacteriosis (BGC) is caused by Campylobacter fetus subsp. venerealis (Cfv) including its biovar intermedius (Cfvi). This sexually transmitted disease induces early reproductive failure causing considerable economic losses in the cattle industry. Using a collection of well-characterized isolates (n = 13), C. fetus field isolates (n = 64) and saprophytic isolates resembling Campylobacter (n = 75) obtained from smegma samples of breeding bulls, this study evaluated the concordance of the most used phenotypic (H2S production in cysteine medium and 1% glycine tolerance) and molecular (PCR) methods for the diagnosis of BGC and assessed possible cross-reactions in the molecular diagnostic methods. Results Characterization at the subspecies level (fetus vs. venerealis) of C. fetus isolated from bull preputial samples using phenotypic and molecular (PCR targeting nahE and ISCfe1) methods showed moderate concordance (κ = 0.462; CI: 0.256–0.669). No cross-reactions were observed with other saprophytic microaerophilic species or with other Campylobacter species that can be present in preputial samples. Whole genome sequencing (WGS) of discrepant isolates showed 100% agreement with PCR identification. For the differentiation of Cfv biovars, comparison of the H2S test (at 72 h and 5 days of incubation) and a PCR targeting the L-cysteine transporter genes showed higher concordance when H2S production was assessed after 5 days (72 h; κ = 0.553, 0.329–0.778 CI vs. 5 days; κ = 0.881, 0.631–1 CI), evidencing the efficacy of a longer incubation time. Conclusions This study confirmed the limitations of biochemical tests to correctly identify C. fetus subspecies and biovars. However, in the case of biovars, when extended incubation times for the H2S test (5 days) were used, phenotypic identification results were significantly improved, although PCR-based methods produced more accurate results. Perfect agreement of WGS with the PCR results and absence of cross-reactions with non-C. fetus saprophytic bacteria from the smegma demonstrated the usefulness of these methods. Nevertheless, the identification of new C. fetus subspecies-specific genes would help to improve BGC diagnosis.https://doi.org/10.1186/s12917-024-03970-8Campylobacter fetus venerealisBiochemical testsPCRWhole Genome Sequencing (WGS)Bovine Genital Campylobacteriosis (BGC) |
| spellingShingle | Nerea Pena-Fernández Nekane Kortabarria Ana Hurtado Medelin Ocejo Marcelo Fort Iratxe Pérez-Cobo Esther Collantes-Fernández Gorka Aduriz Biochemical and molecular characterization of Campylobacter fetus isolates from bulls subjected to bovine genital campylobacteriosis diagnosis in Spain BMC Veterinary Research Campylobacter fetus venerealis Biochemical tests PCR Whole Genome Sequencing (WGS) Bovine Genital Campylobacteriosis (BGC) |
| title | Biochemical and molecular characterization of Campylobacter fetus isolates from bulls subjected to bovine genital campylobacteriosis diagnosis in Spain |
| title_full | Biochemical and molecular characterization of Campylobacter fetus isolates from bulls subjected to bovine genital campylobacteriosis diagnosis in Spain |
| title_fullStr | Biochemical and molecular characterization of Campylobacter fetus isolates from bulls subjected to bovine genital campylobacteriosis diagnosis in Spain |
| title_full_unstemmed | Biochemical and molecular characterization of Campylobacter fetus isolates from bulls subjected to bovine genital campylobacteriosis diagnosis in Spain |
| title_short | Biochemical and molecular characterization of Campylobacter fetus isolates from bulls subjected to bovine genital campylobacteriosis diagnosis in Spain |
| title_sort | biochemical and molecular characterization of campylobacter fetus isolates from bulls subjected to bovine genital campylobacteriosis diagnosis in spain |
| topic | Campylobacter fetus venerealis Biochemical tests PCR Whole Genome Sequencing (WGS) Bovine Genital Campylobacteriosis (BGC) |
| url | https://doi.org/10.1186/s12917-024-03970-8 |
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