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Differential expression of immune response genes associated with subclinical mastitis in dairy buffaloes

  • F. Tanamati (a1), N. B. Stafuzza (a2), D. F. J. Gimenez (a1), A. A. S. Stella (a1), D. J. A. Santos (a1), M. I. T. Ferro (a3), L. G. Albuquerque (a1), E. Gasparino (a4) and H. Tonhati (a1)...

Abstract

Buffalo milk production has become of significant importance on the world scale, however, there are few studies involving biotechnological tools specifically for buffalo. To verify the effects caused by subclinical mastitis on the components of milk and to study the innate immune system in the udder of dairy buffaloes with subclinical mastitis, we evaluated the levels of expression of the lactoferrin (LTF), tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1β), interleukin-8 (IL-8), and toll-like receptors 2 (TLR-2) and 4 (TLR-4) genes in buffaloes with and without subclinical mastitis. Milk samples were collected for the determination of milk components: somatic cell score (SCS), fat, protein, lactose, total solids and solids-not-fat (SNF), as well as for RNA extraction of milk cells, complementary DNA synthesis, and expression profile quantification by quantitative real-time PCR. For gene expression, the ΔΔCt was estimated using contrasts of the target genes expression adjusted for the expression of the housekeeping genes between both groups. Linear regression analysis was performed to determine the relationship between the genes studied and the milk components. Subclinical mastitis induced changes in the fat, lactose and SNF in milk of buffaloes, and the messenger RNA abundance was upregulated for TLR-2, TLR-4, TNF-α, IL-1β and IL-8 genes in milk cells of buffaloes with subclinical mastitis, whereas the LTF gene was not differentially expressed. Results of linear regression analysis showed that TLR-2 gene expression most explains the variation in SCS, and the change in a unit of ΔCt of the TNF-α gene would result in a higher increase in SCS. The study of these immune function genes that are active in the mammary gland is important to characterize the action mechanism of the innate immunity that occurs in subclinical mastitis in dairy buffaloes and may aid the development of strategies to preserve the health of the udder.

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Anand, N, Kanwar, RK, Dubey, ML, Vahishta, RK, Sehgal, R, Verma, AK and Kanwar, JR 2015. Effect of lactoferrin protein on red blood cells and macrophages: mechanism of parasite–host interaction. Drug Design, Development and Therapy 9, 38213835.
Berlutti, F, Schippa, S, Morea, C, Sarli, S, Perfetto, B, Donnarumma, G and Valenti, P 2006. Lactoferrin downregulates pro-inflammatory cytokines upexpressed in intestinal epithelial cells infected with invasive or noninvasive Escherichia coli strains. Biochemistry and Cell Biology 84, 351357.
Carneiro, DMVF, Domingues, PF and Vaz, AK 2009. Imunidade inata da glândula mamária bovina: resposta à infecção. Ciência Rural 39, 19341943.
Dabdoub, SAM and Shook, GE 1984. Phenotypic relations among milk yield, somatic count cells, and mastitis. Journal of Dairy Science 67, 163164.
Dhakal, IP 2006. Normal somatic cell count and subclinical mastitis in Murrah buffaloes. Journal of Veterinary Medicine, Series B 53, 8186.
El-Khodery, SA and Osman, SA 2008. Acute coliform mastitis in buffaloes (Bubalus bubalis): clinical findings and treatment outcomes. Tropical Animal Health and Production 40, 9399.
Fonseca, I, Cardoso, FF, Higa, RH, Giachetto, PF, Brandão, HM, Brito, MAVP, Ferreira, MBD, Guimarães, SEF and Martins, MF 2015a. Gene expression profile in zebu dairy cows (Bos taurus indicus) with mastitis caused by Streptococcus agalactiae. Livestock Science 180, 4757.
Fonseca, LFS, Gimenez, DFJ, Mercadante, MEZ, Bonilha, SFM, Ferro, JA, Baldi, F, Souza, FRP and Albuquerque, LG 2015b. Expression of genes related to mitochondrial function in Nellore cattle divergently ranked on residual feed intake. Molecular Biology Reports 42, 559565.
Fu, Y, Zhou, E, Liu, Z, Li, F, Liang, D, Liu, B, Song, X, Zhao, F, Fen, X, Li, D, Cao, Y, Zhang, X, Zhang, N and Yang, Z 2013. Staphylococcus aureus and Escherichia coli elicit different innate immune responses from bovine mammary epithelial cells. Veterinary Immunology and Immunopathology 155, 245252.
Goldammer, T, Zerbe, H, Molenaar, A, Schuberth, HJ, Brunner, RM, Kata, SR and Seyfert, HM 2004. Mastitis increases mammary mRNA abundance of β-defensin 5, toll-like-receptor 2 (TLR2), and TLR4 but not TLR9 in cattle. Clinical and Diagnostic Laboratory Immunology 11, 174185.
Griesbeck-Zilch, B, Meyer, HHD, Kühn, CH, Schwerin, M and Wellnitz, O 2008. Staphylococcus aureus and Escherichia coli cause deviating expression profiles of cytokines and lactoferrin messenger ribonucleic acid in mammary epithelial cells. Journal of Dairy Science 91, 22152224.
Harmon, RJ 1994. Physiology of mastitis and factors affecting somatic cell counts. Journal of Dairy Science 77, 21032112.
Ibeagha-Awemu, EM, Lee, JW, Ibeagha, AE, Bannerman, DD, Paape, MJ and Zhao, X 2008. Bacterial lipopolysaccharide induces increased expression of toll-like receptor (TLR) 4 and downstream TLR signaling molecules in bovine mammary epithelial cells. Veterinary Research 39, 112.
Kushibiki, S 2011. Tumor necrosis factor-α-induced inflammatory responses in cattle. Animal Science Journal 82, 504511.
Livak, KJ and Schmittgen, TD 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25, 402408.
Medzhitov, R 2007. Recognition of microorganisms and activation of the immune response. Nature 449, 819826.
Moroni, P, Rossi, CS, Pisoni, G, Bronzo, V, Castiglioni, B and Boettcher, PJ 2006. Relationships between somatic cell count and intramammary infection in buffaloes. Journal of Dairy Science 89, 9981003.
Mount, JA, Karrow, NA, Caswell, JL, Boermans, HJ and Leslie, KE 2009. Assessment of bovine mammary chemokine gene expression in response to lipopolysaccharide, lipoteichoic acid+ peptidoglycan and CpG oligodeoxynucleotide 2135. Canadian Journal of Veterinary Research 73, 4957.
Moura, EO, Rangel, AHN, Melo, MCN, Borba, LHF, Lima Júnior, DM, Novaes, LP, Urbano, SA and Andrade Neto, JC 2017. Evaluation of microbiological, cellular and risk factors associated with subclinical mastitis in female buffaloes. Asian-Australasian Journal of Animal Sciences 30, 13401349.
Mukaida, N 2003. Pathophysiological roles of interleukin-8/CXCL8 in pulmonary diseases. American Journal of Physiology. Lung Cellular and Molecular Physiology 284, L566L577.
Oppenheim, JJ, Biragyn, A, Kwak, LW and Yang, D 2003. Roles of antimicrobial peptides such as defensins in innate and adaptive immunity. Annals of the Rheumatic Diseases 62 (suppl. 2), ii17ii21.
Oviedo-Boyso, J, Valdez-Alarcón, JJ, Cajero-Juárez, M, Ochoa-Zarzosa, A, López-Meza, JE, Bravo-Patino, A and Baizabal-Aguirre, VM 2007. Innate immune response of bovine mammary gland to pathogenic bacteria responsible for mastitis. Journal of Infection 54, 399409.
Pasare, C and Medzhitov, R 2004. Toll-like receptors: linking innate and adaptive immunity. Microbes and Infection 6, 13821387.
Petzl, W, Zerbe, H, Günther, J, Yang, W, Seyfert, HM, Nürnberg, G and Schuberth, HJ 2008. Escherichia coli, but not Staphylococcus aureus triggers an early increased expression of factors contributing to the innate immune defense in the udder of the cow. Veterinary Research 39, 123.
Rainard, P and Riollet, C 2006. Innate immunity of the bovine mammary gland. Veterinary Research 37, 369400.
Reyher, KK, Haine, D, Dohoo, IR and Revie, CW 2012. Examining the effect of intramammary infections with minor mastitis pathogens on the acquisition of new intramammary infections with major mastitis pathogens—a systematic review and meta-analysis. Journal of Dairy Science 95, 64836502.
Sarikaya, H, Schlamberger, G, Meyer, HHD and Bruckmaier, RM 2006. Leukocyte populations and mRNA expression of inflammatory factors in quarter milk fractions at different somatic cell score levels in dairy cows. Journal of Dairy Science 89, 24792486.
Sharma, BS, Leyva, I, Schenkel, F and Karrow, NA 2006. Association of toll-like receptor 4 polymorphisms with somatic cell score and lactation persistency in Holstein bulls. Journal of Dairy Science 89, 36263635.
Sordillo, LM and Streicher, KL 2002. Mammary gland immunity and mastitis susceptibility. Journal of Mammary Gland Biology and Neoplasia 7, 135146.
Steibel, JP, Poletto, R, Coussens, PM and Rosa, GJ 2009. A powerful and flexible linear mixed model framework for the analysis of relative quantification RT-PCR data. Genomics 94, 146152.
Tonhati, H, Cerón-Muñoz, MF, Oliveira, JA, Duarte, JMC, Furtado, TP and Tseimazides, SP 2000. Parâmetros genéticos para produção de leite, gordura e proteína em bubalinos. Revista Brasileira de Zootecnia 29, 20512056.
Vandesompele, J, De Preter, K, Pattyn, F, Poppe, B, Van Roy, N, De Paepe, A and Speleman, F 2002. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biology 3, research0034.1research0034.12.
Wellnitz, O and Bruckmaier, RM 2012. The innate immune response of the bovine mammary gland to bacterial infection. The Veterinary Journal 192, 148152.
Yu, L, Wang, L and Chen, S 2010. Endogenous toll-like receptor ligands and their biological significance. Journal of Cellular and Molecular Medicine 14, 25922603.
Zähringer, U, Lindner, B, Inamura, S, Heine, H and Alexander, C 2008. TLR2–promiscuous or specific? A critical re-evaluation of a receptor expressing apparent broad specificity. Immunobiology 213, 205224.
Zhu, YH, Liu, PQ, Weng, XG, Zhuge, ZY, Zhang, R, Ma, JL, Qiu, XQ, Li, RQ, Zhang, XL and Wang, JF 2012. Pheromonicin-SA affects mRNA expression of toll-like receptors, cytokines, and lactoferrin by Staphylococcus aureus-infected bovine mammary epithelial cells. Journal of Dairy Science 95, 759764.

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