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Development of a flow cytometric assay to assess the bacterial count in boar semen


Oehler, Christin; Janett, Fredi; Schmitt, Sarah; Malama, Eleni; Bollwein, Heiner (2019). Development of a flow cytometric assay to assess the bacterial count in boar semen. Theriogenology, 133:125-134.

Abstract

The aim of the study was to develop a new flow cytometric assay for the determination of the bacterial count in commercially processed boar semen. In total 224 fresh boar semen samples collected at an AI-station were analyzed. The number of total viable counts (TVC) was determined by using flow cytometry after staining with SYBR Green I and Propidium Iodide (PI). In the first part of the study 111 fresh boar semen samples were spiked with pure cultures of defined numbers of bacteria commonly detected in boar ejaculates and analyzed by flow cytometry. In the second part, 113 fresh semen samples were assessed on the day of collection through flow cytometry and the Most Probable Number (MPN) method, as the standard bacteriological method. The first part of the study showed a strong correlation between the detected and expected numbers (r = 0.96; P < 0.001), while in the second part of the study the TVC determined by flow cytometry and by the MPN method correlated only moderately (r = 0.28; P < 0.01; median MPN: 24,000 ± MAD 21,600 bacteria/mL; median flow cytometry: 24,426 ± MAD 15,610 bacteria/mL). In summary flow cytometry is a fast alternative to the classical culture technique to determine highly contaminated boar ejaculates. The developed flow cytometric protocol enables one to enumerate the viable bacteria within fresh boar ejaculates without requiring numerous treatment steps, and thus offering the possibility of an on-line use in AI-centers.

Abstract

The aim of the study was to develop a new flow cytometric assay for the determination of the bacterial count in commercially processed boar semen. In total 224 fresh boar semen samples collected at an AI-station were analyzed. The number of total viable counts (TVC) was determined by using flow cytometry after staining with SYBR Green I and Propidium Iodide (PI). In the first part of the study 111 fresh boar semen samples were spiked with pure cultures of defined numbers of bacteria commonly detected in boar ejaculates and analyzed by flow cytometry. In the second part, 113 fresh semen samples were assessed on the day of collection through flow cytometry and the Most Probable Number (MPN) method, as the standard bacteriological method. The first part of the study showed a strong correlation between the detected and expected numbers (r = 0.96; P < 0.001), while in the second part of the study the TVC determined by flow cytometry and by the MPN method correlated only moderately (r = 0.28; P < 0.01; median MPN: 24,000 ± MAD 21,600 bacteria/mL; median flow cytometry: 24,426 ± MAD 15,610 bacteria/mL). In summary flow cytometry is a fast alternative to the classical culture technique to determine highly contaminated boar ejaculates. The developed flow cytometric protocol enables one to enumerate the viable bacteria within fresh boar ejaculates without requiring numerous treatment steps, and thus offering the possibility of an on-line use in AI-centers.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:05 Vetsuisse Faculty > Institute of Food Safety and Hygiene
05 Vetsuisse Faculty > Veterinary Clinic > Department of Farm Animals
Dewey Decimal Classification:570 Life sciences; biology
630 Agriculture
Uncontrolled Keywords:Food Animals, Animal Science and Zoology, Equine, Small Animals, Bacteria; Boar; Flow cytometry; Semen
Language:English
Date:1 July 2019
Deposited On:16 Dec 2019 11:58
Last Modified:29 Feb 2020 08:25
Publisher:Elsevier
ISSN:0093-691X
OA Status:Closed
Publisher DOI:https://doi.org/10.1016/j.theriogenology.2019.04.036
PubMed ID:31091483

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