Balb/c farelerinde meme içi lipopolisakkarid uygulaması sonucu yeni bir kronik mastitis modelin indüklenmesi

Abstract

Mastitis, meme dokusunun iltihaplanmasıdır ve dünya genelinde çiftlik hayvanlarında yaygın olarak görülmektedir. Bu sorun süt endüstrisinde veteriner masrafları, sütün imhası ve tedavi giderleri açısından ciddi mali kayıplara neden olmaktadır. Bakteriler etiyolojinin ana aktörleridir ve meme dokusunda akut ve kronik inflamatuar değişikliklere neden olurlar. Akut enflamatuar değişiklikler klinik olarak kolayca tanınır ve tedavi hemen başlatılır, ancak subakut inflamasyon sinsice ilerler ve geri dönüşü olmayan fibrotik değişikliklerle kronik inflamasyona yol açar. Laboratuvar hayvanlarında akut mastitis indüksiyonu için standartlaştırılmış deneysel modeller mevcuttur ve genellikle bakterilerin veya bazı bakteriyel yapısal bileşenlerin meme dokusuna infüzyonu bu amaçla kolayca uygulanmaktadır. Bununla birlikte, fibrotik değişikliklerle birlikte kronik mastitis indüksiyonu üzerine az sayıda çalışma vardır ve uygulamalar nispeten karmaşıktır. Sunulan çalışmada, farelerde kronik mastitisi indüklemek için LPS 0, 5 ve 10. günlerde üç kez meme kanalından infüze edilmiştir. Histopatolojik değişiklikleri değerlendirmek için 1, 6 ve 15. günlerde dokulardan örnek alınmıştır. Akut inflamasyonun bir bileşeni olan şiddetli nötrofil infiltratları 1. günde gözlenirken, lenfosit infiltratları subakut inflamasyonla uyumlu olarak 6. günde artmıştır. 15. günde, fibrozis ve lenfosit infiltrasyonu gibi kronik mastitisi temsil eden lezyonlar gözlenmiştir. Süt sığırlarının kronik mastitisindeki lezyonlara benzer bir model, farelerde LPS infüzyonu ile başarılı ve kolay bir şekilde oluşturulmuştur.

Keywords

• Balb/c
• Kronik mastitis modeli
• Lipopolisakkarit
• Meme bezi
• Masson Trikrom

Project Number

118O893

A new protocol for the induction of chronic mastitis with intramammary infusion of lipopolysaccharide (LPS) in Balb/c mice

Abstract

Mastitis is inflammation of the mammary tissue and is commonly observed in farm animals. The problem causes severe financial losses in the dairy industry in terms of veterinary costs, milk disposal, and treatment expenses. Bacteria are the main actors in the etiology and cause acute and chronic inflammatory changes in the mammary tissue. Acute inflammatory changes are easily recognized clinically, and treatment is initiated immediately, but subacute inflammation progresses insidiously and leads to chronic inflammation with irreversible fibrotic changes. Standardized experimental models for the induction of acute mastitis in laboratory animals are available. Usually, infusion of bacteria or some bacterial structural components into mammary tissue is easily applied for this purpose. However, there are few studies on the induction of chronic mastitis with fibrotic changes, and the applications are relatively complex. In this study, LPS was infused through the teat duct three times on days 0, 5, and 10 to induce chronic mastitis in mice. Tissues were sampled on days 1, 6, and 15 to evaluate histopathological changes. While severe neutrophil infiltrates, a component of acute inflammation, were observed on day 1, lymphocyte infiltrates increased on day 6, consistent with subacute inflammation. On day 15, lesions representing chronic mastitis, such as fibrosis and lymphocyte infiltration, were observed. A model similar to the lesions in chronic mastitis of dairy cattle was successfully and easily established by LPS infusion in mice.

Keywords

• Balb/c
• Chronic mastitis model
• Lipopolysaccharide
• Mammary gland
• Masson’s Trichrome

Supporting Institution

TÜBİTAK

Project Number

118O893

Ethical Statement

This study was approved by the Bursa Uludag University Animal Experiments Local Ethics Committee (2019/07-04 Number Ethics Committee Decision). In addition, the authors declared that Research and Publication Ethical rules were followed.

References

1. Anderson JC, 1979: The epidemiology and pathogenesis of experimental Staphylococcal and coliform mastitis in the mouse. Br Vet J, 135(2), 163-171. doi:10.1016/s0007-1935(17)32937-8.
2. Barham W, Sherrill T, Connelly L, Blackwell TS, Yull FE, 2012: Intraductal injection of LPS as a mouse model of mastitis: signaling visualized via an NF-κB reporter transgenic. J Vis Exp: JoVE, (67), e4030. https://doi.org/10.3791/4030
3. Bi Y, Ding Y, Wu J, Miao Z, Wang J, Wang F, 2020: Staphylococcus aureus induces mammary gland fibrosis through activating the TLR/NF-κB and TLR/AP-1 signaling pathways in mice. Microb Pathog, 148:104427. doi: 10.1016/j.micpath.2020.104427
4. Brouillette E, Millette G, Chamberland S, Roy JP, Ster C, Kiros T, Hickey S, Hittle L, Woolston J, Malouin F, 2023: Effective Treatment of Staphylococcus aureus Intramammary Infection in a Murine Model Using the Bacteriophage Cocktail StaphLyse™. Viruses, 15(4), 887. https://doi.org/10.3390/v15040887
5. Camperio C, Armas F, Biasibetti E., Frassanito P, Giovannelli C, Spuria L, D’Agostino C, Tait S, Capucchio MT, Marianelli C, 2017: A mouse mastitis model to study the effects of the intramammary infusion of a food-grade Lactococcus lactis strain. PLoS One, 12(9), e0184218. doi: 10.1371/journal.pone.0184218
6. Cheng WN, Han SG, 2020: Bovine mastitis: risk factors, therapeutic strategies, and alternative treatments - A review. AJAS, 33(11), 1699-1713. doi:10.5713/ajas.20.0156
7. Cobirka, M, Tancin V, Slama P, 2020: Epidemiology and classification of mastitis. Animals, 10(12), 2212. doi:10.3390/ani10122212
8. Ingman WV, Glynn DJ, Hutchinson MR, 2014: Inflammatory mediators in mastitis and lactation insufficiency. J Mammary Gland Biol and Neoplasia, 19(2), 161-167. doi:10.1007/s10911-014-9325-9

9. Ingman WV, Glynn DJ, Hutchinson MR, 2015: Mouse models of mastitis - how physiological are they?. Int Breastfeed J, 10, 12. doi:10.1186/s13006-015-0038-5
10. Kan X, Hu G, Liu Y, Xu P, Huang Y, Cai X, Guo W, Fu S, Liu J, 2022: Mammary fibrosis tendency and mitochondrial adaptability in dairy cows with mastitis. Metabolites, 12(11), 1035. doi.org/10.3390/metabo12111035
11. Lai JL, Liu YH, Peng YC, Ge P, He CF, Liu C, Chen YY, Guo AZ, Hu CM, 2017: Indirubin treatment of lipopolysaccharide-induced mastitis in a mouse model and activity in mouse mammary epithelial cells. Mediators of Inflamm, doi:10.1155/2017/3082805
12. Özgüden-Akkoc CG, Mutlu AM, Keskin A, Yumuşak E, Akkoc A, 2023: Phenotypic evaluation of mast cells in bovine mammary tissue and mastitis in the context of fibrosis. JDS, 90(4), 387–392. https://doi.org/10.1017/S0022029923000651
13. Rollin E, Dhuyvetter KC, Overton MW, 2015: The cost of clinical mastitis in the first 30 days of lactation: An economic modeling tool. Prev Vet Med, 122(3), 257-264. doi: 10.1016/j.prevetmed.2015.11.006
14. Tuchscherr, LP, Buzzola FR, Alvarez LP, Caccuri RL, Lee JC, Sordelli DO, 2005: Capsule-negative Staphylococcus aureus induces chronic experimental mastitis in mice. Infect Immun, 73(12), 7932-7937. doi:10.1128/IAI.73.12.7932-7937.2005
15. Wynn TA, Ramalingam TR 2012: Mechanisms of fibrosis: therapeutic translation for fibrotic disease. Nat Med, 18(7), 1028-1040. doi:10.1038/nm.2807
16. Wynn TA, 2008: Cellular and molecular mechanisms of fibrosis. J Pathol, 214(2), 199-210. doi:10.1002/path.2277
17. Zhao XW, Yang YX, Huang DW, Cheng GL, Zhao HL, 2015: Comparative proteomic analysis of proteins expression changes in the mammary tissue of c ows infected with Escherichia coli mastitis. J Vet Sci, 16(3), 253-263. doi:10.4142/jvs.2015.16.3.253