Mammary immunity of White Park and Highland cattle compared with Brown Swiss and Red Holstein

Abstract

Mastitis is a frequent disease in modern dairy cows, but ancient cattle breeds seem to be naturally more resistant to it. Primary bovine mammary epithelial cells from the ancient Highland and White Park (n = 5) cattle and the modern dairy breeds Brown Swiss and Red Holstein (n = 6) were non-invasively isolated from milk, cultured, and stimulated with the heat-inactivated mastitis pathogens Escherichia coli and Staphylococcus aureus to compare the innate immune response in vitro. With reverse transcription quantitative polymerase chain reaction (RT-qPCR), the breeds differed in the basal expression of 16 genes. Notably CASP8, CXCL8, Toll-like receptors 2 and 4 (TLR2 and TLR4) expression were higher in the ancient breeds (P < 0.05). In the modern breeds, more genes were regulated after stimulation. Breed differences (P < 0.05) were detected in C3, CASP8, CCL2, CD14, LY96 and transforming growth factor β1 (TGFβ1) regulation. Principal component analysis separated the ancient from the modern breeds in their basal expression, but not after stimulation. ELISA of lactoferrin and serum amyloid A protein revealed breed differences in control and S. aureus treated levels. The immune reaction of ancient breeds seemed less intensive because of a higher basal expression, which has been shown before to be beneficial for the animal. For the first time, the innate immune response of these ancient breeds was studied. Previous evidence of breed and animal variation in innate immunity was confirmed. Keywords: breed comparison, primary bovine mammary epithelial cells, innate immune response, ancient and modern cattle breeds, mastitisRésumé La mastite est une maladie fréquente chez les vaches laitières modernes. Or, les races bovines anciennes semblent être naturellement plus résistantes. Dans le présent travail, des cellules primaires bovines épithéliales mammaires des races anciennes Highland et White Park (n = 5), ainsi que des races laitières modernes Brown Swiss et Red Holstein (n = 6) ont été isolées du lait de façon non-invasive. Ensuite, elles ont été cultivées, puis stimulées avec les pathogènes de la mastite Escherichia coli et Staphylocoque dorétous les deux préalablement inactivés par la chaleurpour ainsi comparer la réponse immunitaire innée in vitro, utilisant la technique reverse transcription quantitative polymerase chain reaction (RT-qPCR). Il s'avère que les races diffèrent dans l'expression basale de 16 gènes. Notamment, les expressions de CASP8, CXCL8, TLR2 et TLR4 étaient élevées dans les races anciennes (P < 0.05). Dans les races modernes, c'est le nombre global des gènes régulés après stimulation qui était plus élevé. Des différences entre les races (P < 0.05) ont été détectées quant à la régulation de C3, CASP8, CCL2, CD14, LY96 et TGFβ1. L'analyse des composantes principales a permis de cloisonner les races anciennes des races modernes dans l'expression basale, mais pas après stimulation. Les mesures ELISA de lactoferrin et de sérum amyloïde A protéine ont dévoilé des différences interraciales entre le groupe...

Effect of the Ketone Body Beta-Hydroxybutyrate on the Innate Defense Capability of Primary Bovine Mammary Epithelial Cells

“…coli . This is in accordance with published data [ 12 ], where E . coli also caused a strong induction of the gene expression of those defense molecules.…”

“…Cows with a somatic cell count (SCC) below 200,000 cells per milliliter milk, were considered healthy. The cell isolation was basically conducted as described in previous studies [ 12 , 13 ], with slight changes in the washing procedure. In brief, milk cells were harvested (10 min, 1850 × g), the cell pellet was washed with 1x HBSS buffer (Sigma-Aldrich, Saint Louis, USA) supplemented with antibiotics and antimycotics (penicillin/streptomycin, amphotericin B, Sigma-Aldrich) and the cell suspension was filtered twice through filters with different pore sizes (100 μm, 40 μm, Greiner Bio-One GmbH, Frickenhausen, Germany).…”

“…pbMEC represent the first cellular line of defense, after a pathogen manages to cross the teat barrier, therefore pbMEC express transepithelial receptors, like the toll-like receptors (TLR) on their cell surface [ 10 ]. Through the activation of the TLR-pathway by pathogen associated molecular patterns, pbMEC are able to induce a downstream signaling cascade, resulting in the activation of the transcription factor NFĸB and hence in the production of chemotactic molecules like pro-inflammatory chemokines, cytokines and acute phase proteins [ 11 , 12 ]. Thus, the recruitment of other immune cells, like leucocytes and macrophages to the site of infection is one of the main tasks of pbMEC in innate immunity.…”

Gene expression profiling in pbMEC – in search of molecular biomarkers to predict immunoglobulin production in bovine milk

“…The pbMEC were isolated from fresh milk of nine healthy Brown Swiss cows in mid-lactation, as described by Sorg et al (2013a) and Danowski et al (2013) [ 14 , 15 ]. In brief, fresh milk was defatted (10 min, 1850× g ), and the resulting cell pellet was washed several times with 1× Hanks Balanced Salt solution (Sigma-Aldrich) supplemented with antibiotics and antimycotics [ 14 , 15 ]. The remaining cell suspension was filtered twice (EASYstrainer™; 40 μm; 100 μm; Greiner Bio-One GmbH, Frickenhausen, Germany) to remove lipid droplets and cell aggregates.…”

Natural resistance of the Ukrainian gray cows

“…Втрата біо різно ма ніт тя на сучасному етапі спричинила роз роб лен ня спільних заходів світового спів това рис тва щодо збереження племінних ре сур сів [1, 2]. У розвинутих країнах знач ний ін те рес ви яв ляють до всебічного до слід ження ло каль них та автохтонних по рід сільсько госпо дар ських тварин, які відрізняються від по ши ре них ко мер ційних високим ступенем при сто су ван ня до місцевих умов, кращою при род ною ре зис тентністю та іншими цінними якос тя ми [3,4,5,6]. Ратифікація Україною Гло баль ного плану дій в області генетичних ре сур сів тварин, Інтерлакенської декларації та Нагой ського протоколу спонукає до розроб лен ня і вжиття заходів для збереження наяв но го біо різ но маніття.…”