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

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 postinoculation; 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\u2014this was significantly greater during the first 24 h postchallenge (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 \u201cpositive\u201d / \u201cnegative\u201d assessment would be sufficient

A detailed questionnaire for the evaluation of health management in dairy sheep and goats

The objective of this work was to develop, use and present a detailed questionnaire for the evaluation of health management in dairy small ruminants; it includes 442 questions organised in seven sections: general, infrastructure, animals, production characteristics, health management, nutrition, human resources. Consistency of replies was evaluated in 27 farmers, interviewed twice. Inconsistent replies were given by all farmers to 30 different questions (Cronbach’s coefficient alpha: 0.987). Then, interviews were performed in 444 farms around Greece. Mean duration of an interview was 63.6 min. Clarifications were requested by 273 farmers to 22 different questions (maximum per farmer: 8). The experience of the investigator, the primary language of farmers and asking clarifications by the farmers affected the duration of the interview. The questionnaire can be used for research work in the field, to record details in the farms under study. In accord with the needs of a particular study, it can be modified, by adding more specific questions or omitting others deemed of less importance. Moreover, it can also be used for routine monitoring purposes, as a useful means to record and maintain details of farms during clinical work. To the best of our knowledge, the questionnaire is the most extensive and detailed one available internationally for dairy small ruminants

Use of proteomics in the study of mastitis in ewes

The objective of this review is to describe the usage and applicability of proteomics technologies in the study of mastitis in ewes. In ewes, proteomics technologies have been employed for furthering knowledge in mastitis caused by various agents (Staphylococcus aureus, Staphylococcus chromogenes, Mannheimia haemolytica, Streptococcus uberis, Mycoplasma agalactiae). Studies have focused on improving knowledge regarding pathogenesis of the infections and identifying biomarkers for its diagnosis. Findings have revealed that ewes with mastitis mount a defence response, controlled by many proteins and over various mechanisms and pathways, which are interdependent at various points. Many proteins can participate in this process. Moreover, as the result of proteomics studies, cathelicidins and serum amyloid A have been identified as proteins that can be used as biomarkers for improved diagnosis of the disease. In the long term, proteomics will contribute to improvements in the elucidation of the pathogenesis of mastitis. Further in-depth investigations into the various proteomes and application of new methodological strategies in experimental and clinical studies will provide information about mastitis processes, which will be of benefit in controlling the disease. Improvement of diagnostic techniques, establishment of prognostic tools and development of vaccines are key areas for targeted research. © 2019 by the authors. Licensee MDPI, Basel, Switzerland

Applied proteomics in companion animal medicine

Background: Proteomics in companion animal medicine has been used chiefly, in order to identify proteins, which may be used as biomarkers for early diagnosis of a variety of pathological conditions, as well as to elucidate pathogenesis of various diseases by describing, at molecular level, signal transductions in diseased organs. Objective: The review of some of currently available knowledge in proteomics, as can be applied in companion animal medicine and can be of use to veterinarians active in the field, in order to solve scientific or clinical questions regarding signal transduction. Method: Specific clinical applications of the methodologies are reviewed. Results: Proteomics may be employed in supporting early and accurate diagnosis of leishmaniosis, as well as for the identification of proteins of the causative protozoon, and can also be helpful in cases of neoplastic diseases; proteins identified in blood serum or tears of dogs or cats may be used to support diagnosis of various neoplastic conditions and to monitor treatment and predict future metastases. Proteomics have helped to identify proteins with diagnostic significance in the blood serum of Cavalier King Charles Spaniels with myxomatous mitral valve degeneration and in the bronchoalveolar lavage fluid of West Highland White Terriers with idiopathic pulmonary fibrosis and have also been used for studying the musculoskeletal system of horses, specifically in laminitis, osteoarthritis and osteochondrosis and exertional rhabdomyolysis. The technology has also been applied in various other diseases of dogs and cats (e.g., Leptospira infections, Babesia infections, Golden Retriever muscular dystrophy, degenerative myelopathy, degenerative changes in the brain, azotaemia, periodontitis), as well as in uveitis and endometritis in horses and in staphylococcal infections and tularaemia in rabbits. Conclusion: In the long term, proteomics will contribute to improvements in all facets of companion animal medicine. Mining deeper into the various proteomes and application of new methodological strategies in clinical studies will provide information about disease processes, which will be of benefit to practising veterinarians. Improvement of diagnostic techniques, establishment of prognostic tools and development of vaccines against diseases are key-areas for targeted research. � 2016 Bentham Science Publishers

Frequency and severity of enzootic pneumonia-like lesions in Greek swine herds and their association with different vaccination protocols against Mycoplasma hyopneumoniae

Several European countries, but Greece, reported data of Enzootic Pneumonia (EP)-like lesions at slaughter. In the present study, frequency and severity of EP-like lesions were evaluated at slaughter and their association with different vaccination protocols for Mycoplasma hyopneumoniae, the primary pathogen of EP, was investigated, in Greek swine herds. In total, 7047 lungs from 53 farrow-to-finish herds were scored at slaughter by using the Ceva Lung ProgramTM tool, from January 2016 to December 2019. The frequency of EP-like lesions was 47.6% and the median (range) of the total lung lesions score was 0 (0–25). Pigs, which were vaccinated with a monovalent vaccine for M. hyopneumoniae either once or twice and those which received an extra shot, were less likely to have EP-like lesions, compared to pigs that received a bivalent vaccine containing both M. hyopneumoniae and PCV2 antigens. For the pigs which received two shots of a monovalent vaccine, the odds of higher versus lower lung lesion score(s) were lower compared to pigs that received a bivalent vaccine. Almost half of the lungs were affected, although 98.3% of the pigs were vaccinated. Poor housing and management, commonly seen in Greek herds, may hamper the efficient reduction of M. hyopneumoniae spread. © 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group

Anti-staphylococcal biofilm antibodies in ewes and association with subclinical mastitis

Objective of the present work was the investigation of anti-poly-N-acetyl b-1,6 glucosamine (PNAG) staphylococcal antibodies in ewes and the study of factors that may be associated with these. During a field investigation in subclinical mastitis in Greece, milk samples for bacteriological and cytological examination for detection of staphylococcal mastitis and blood samples for detection of anti-PNAG antibodies were collected from 355 ewes in 71 flocks. Prevalence of seropositivity was higher among ewes vaccinated against staphylococcal mastitis (0.750) than non-vaccinated ones (0.458). Mean antibody titres in samples from vaccinated ewes (13.64) were significantly higher than in samples from non-vaccinated ones (7.39). There was no association between identity of staphylococcal isolates recovered from milk samples and antibody titres in blood samples from respective ewes. Among vaccinated ewes, there was correlation (r = 0.922) between total number of genes relevant to biofilm-formation in mastitis staphylococcal isolates and antibody titres in blood samples from respective ewes; further, isolates from ewes with seropositive samples had higher median number of genes (4) than isolates from ewes with seronegative samples (3). In a multivariable mixed-effects model, vaccination emerged as the only significant factor for the prevalence of seropositivity of anti-PNAG antibody titres; when only unvaccinated ewes were included in the model, no factor was found of significance for that prevalence. © 2019 Elsevier B.V

Applied proteomics in ‘one health’

‘One Health’ summarises the idea that human health and animal health are interdependent and bound to the health of ecosystems. The purpose of proteomics methodologies and studies is to determine proteins present in samples of interest and to quantify changes in protein expression during pathological conditions. The objectives of this paper are to review the application of proteomics technologies within the One Health concept and to appraise their role in the elucidation of diseases and situations relevant to One Health. The paper develops in three sections. Proteomics Applications in Zoonotic Infections part discusses proteomics applications in zoonotic infections and explores the use of proteomics for studying pathogenetic pathways, transmission dynamics, diagnostic biomarkers and novel vaccines in prion, viral, bacterial, protozoan and metazoan zoonotic infections. Proteomics Applications in Antibiotic Resistance part discusses proteomics applications in mechanisms of resistance development and discovery of novel treatments for antibiotic resistance. Proteomics Applications in Food Safety part discusses the detection of allergens, exposure of adulteration, identification of pathogens and toxins, study of product traits and characterisation of proteins in food safety. Sensitive analysis of proteins, including low-abundant ones in complex biological samples, will be achieved in the future, thus enabling implementation of targeted proteomics in clinical settings, shedding light on biomarker research and promoting the One Health concept. © 2021 by the authors. Licensee MDPI, Basel, Switzerland

Proteomics analysis in dairy products: Cheese, a review

Cheese is a worldwide produced and consumed commodity. There are many varieties of cheese from soft to hard, white to yellow, and fresh to aged after ripening. Especially, each category has its own producing technology. Many countries have labeled their most traditional cheese as Protective Designation of Origin (PDO). Moreover, several studies using advanced technologies, such as proteomics, have been performed to enhance labeling. In this review, broadly diffused and marketed, as well as Mediterranean countries’ special interest in Mediterranean diet-related PDO cheeses have been chosen as a reference. The aim of this work was to highlight the use of proteomics methods to examine how cheese proteins and peptides rearrange after ripening and use of starters. Further, we aimed to examine what kind of proteins are produced. Finally, we focused on bioactive molecules in cheeses and distinction of the original product from its counterfeit. © 2021 by the authors. Licensee MDPI, Basel, Switzerland

Mammary defences and immunity against mastitis in sheep

The objectives of this review paper are to present udder defences, including teat of the udder, mammary epithelial cells, leucocytes, immunoglobulins, complement systemand chemical antibacterial agents, to describe cooperation and interactions between them and to elaborate on potentials regarding their significance inmammary immunisation strategies. The teat of the udder provides initial protection to the mammary gland. The mammary epithelial cells synthesise antibacterial proteins and the leucocytes produce various inflammation mediators (cytokines or chemokines), phagocytose bacteria and recognise antigenic structures. In themammary gland, four immunoglobulins (IgG1, IgG2, IgMand IgA) have important roles against bacterial pathogens. The complement systemis a collection of proteins, participating in the inflammatory process through various pathways. Other components contributing to humoral mammary defence include lactoferrin, lysozyme and the lactoperoxidase/myeloperoxidase systems, as well as oligosaccharides, gangliosides, reactive oxygen species, acute phase proteins (e.g., haptoglobin and serum amyloid A), ribonucleases and a wide range of antimicrobial peptides. Management practices, genetic variations and nutrition can influence mammary defences and should be taken into account in the formulation of prevention strategies against ovine mastitis. © 2019 by the authors. Licensee MDPI, Basel, Switzerland

Interactions between parasitism and milk production - Mastitis in sheep

The present review discusses interactions and focuses in the direct effects of parasites in production of milk and development of mastitis in ewes. In most relevant published studies, a clear reduction in milk yield of ewes affected with parasites has been recorded. This reduction in milk yield has been quantified to average 22% among the relevant studies. Nematode parasitism can also cause shorter persistency of lactation, whilst the situation regarding potential effects on milk composition is not clear. Moreover, in field and experimental studies trichostrongylid intestinal infections have been documented to increase risk of mastitis in affected ewes; among parasitic species, the effect was associated primarily with Teladorsagia spp. infection. Trematode infection has also been found to increase risk for mastitis development at the early stage of lactation period. Research supports that effective parasite control will enable sheep to fully express their genetic potential and benefit for increased milk yield and effective udder health management schemes. © 2019 Elsevier B.V