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Detection of Cathelicidin-1 in the Milk as an Early Indicator of Mastitis in Ewes

1
Veterinary Faculty, University of Thessaly, 43100 Karditsa, Greece
2
Proteomics Research Unit, Biomedical Research Foundation of Academy of Athens, 11527 Athens, Greece
3
Laboratory of Parasitology and Parasitic Diseases, Faculty of Veterinary Medicine, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece
4
Dipartimento di Medicina Veterinaria, Università degli Studi di Milano, Via G. Celoria 10, 20133 Milan, Italy
*
Author to whom correspondence should be addressed.
Pathogens 2019, 8(4), 270; https://doi.org/10.3390/pathogens8040270
Received: 22 October 2019 / Revised: 26 November 2019 / Accepted: 27 November 2019 / Published: 28 November 2019
(This article belongs to the Special Issue Mastitis in Dairy Ruminants)
The objective of the study was the investigation of the behaviour of cathelicidin-1 in the milk after experimental infection with two prominent bacterial pathogens (experiment 1: Mannheimia haemolytica, experiment 2: M. haemolytica and Staphylococcus chromogenes) as a potential early indicator for diagnosis of mastitis in sheep. In two experiments, after bacterial inoculation into the udder of ewes, bacteriological and cytological examinations of milk samples as well as proteomics examinations [two-dimensional gel electrophoresis analysis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF MS) analysis] were performed sequentially. Cathelicidin-1 was detected and spot densities obtained from PDQuest v.8.0 were recorded. Associations were calculated between cell content and spot densities as well as between presence of mastitis in a mammary gland at a given time-point and detection of cathelicidin-1 in the respective milk sample. All inoculated mammary glands developed mastitis, confirmed by the consistent bacterial isolation from mammary secretion and increased leucocyte content therein. Spot density of cathelicidin-1 in samples from inoculated glands increased 3 h post-inoculation; spot density of cathelicidin-1 in samples from inoculated glands was higher than in samples from uninoculated controls. There was clear evidence of correlation between cell content and cathelicidin-1 spot densities in milk samples. There was significant association between presence of mastitis in the mammary gland and detection of cathelicidin-1 in the respective milk sample; overall accuracy was 0.818—this was significantly greater during the first 24 h post-challenge (0.903) than after the first day (0.704). In conclusion, detection of cathelicidin-1 in milk was significantly associated with presence of mastitis in ewes. The associations were stronger during the first 24 h post-infection than after the first day. Cathelicidin-1 has the advantage that it can be a non-specific biomarker, as simply a “positive”/“negative” assessment would be sufficient. View Full-Text
Keywords: biomarker; cathelicidin-1; diagnosis; mastitis; proteomics; sheep; somatic cell counts biomarker; cathelicidin-1; diagnosis; mastitis; proteomics; sheep; somatic cell counts
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Katsafadou, A.I.; Tsangaris, G.T.; Vasileiou, N.G.; Ioannidi, K.S.; Anagnostopoulos, A.K.; Billinis, C.; Fragkou, I.A.; Papadopoulos, E.; Mavrogianni, V.S.; Michael, C.K.; Addis, M.F.; Fthenakis, G.C. Detection of Cathelicidin-1 in the Milk as an Early Indicator of Mastitis in Ewes. Pathogens 2019, 8, 270.

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