Hostname: page-component-cd9895bd7-q99xh Total loading time: 0 Render date: 2024-12-22T18:48:21.128Z Has data issue: false hasContentIssue false
  • English
  • Français

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

Published online by Cambridge University Press:  09 January 2019

F. Tanamati Open the ORCID record for F. Tanamati [Opens in a new window] ,
N. B. Stafuzza ,
D. F. J. Gimenez ,
A. A. S. Stella ,
D. J. A. Santos ,
M. I. T. Ferro ,
L. G. Albuquerque ,
E. Gasparino  and
H. Tonhati
F. Tanamati*
Affiliation:
Department of Animal Science, São Paulo State University (FCAV/UNESP), Jaboticabal, SP 14884-900, Brazil
N. B. Stafuzza
Affiliation:
Department of Exact Sciences, São Paulo State University (FCAV/UNESP), Jaboticabal, SP 14884-900, Brazil
D. F. J. Gimenez
Affiliation:
Department of Animal Science, São Paulo State University (FCAV/UNESP), Jaboticabal, SP 14884-900, Brazil
A. A. S. Stella
Affiliation:
Department of Animal Science, São Paulo State University (FCAV/UNESP), Jaboticabal, SP 14884-900, Brazil
D. J. A. Santos
Affiliation:
Department of Animal Science, São Paulo State University (FCAV/UNESP), Jaboticabal, SP 14884-900, Brazil
M. I. T. Ferro
Affiliation:
Department of Technology, São Paulo State University (FCAV/UNESP), Jaboticabal, SP 14884-900, Brazil
L. G. Albuquerque
Affiliation:
Department of Animal Science, São Paulo State University (FCAV/UNESP), Jaboticabal, SP 14884-900, Brazil
E. Gasparino
Affiliation:
Department of Animal Science, Maringá State University (UEM), Maringá, PR 87020-900, Brazil
H. Tonhati
Affiliation:
Department of Animal Science, São Paulo State University (FCAV/UNESP), Jaboticabal, SP 14884-900, Brazil
*
†E-mail: [email protected]
Get access

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.

Keywords

Bubalus bubalisinnate immunitymRNAquantitative PCRsomatic cells
Type
Research Article
Information
animal , Volume 13 , Issue 8 , August 2019 , pp. 1651 - 1657
Copyright
© The Animal Consortium 2019 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

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, 3821–3835.Google Scholar
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, 351–357.Google Scholar
Carneiro, DMVF, Domingues, PF and Vaz, AK 2009. Imunidade inata da glândula mamária bovina: resposta à infecção. Ciência Rural 39, 1934–1943.Google Scholar
Dabdoub, SAM and Shook, GE 1984. Phenotypic relations among milk yield, somatic count cells, and mastitis. Journal of Dairy Science 67, 163–164.Google Scholar
Dhakal, IP 2006. Normal somatic cell count and subclinical mastitis in Murrah buffaloes. Journal of Veterinary Medicine, Series B 53, 81–86.Google Scholar
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, 93–99.Google Scholar
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, 47–57.Google Scholar
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, 559–565.Google Scholar
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, 245–252.Google Scholar
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, 174–185.Google Scholar
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, 2215–2224.Google Scholar
Harmon, RJ 1994. Physiology of mastitis and factors affecting somatic cell counts. Journal of Dairy Science 77, 2103–2112.Google Scholar
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, 1–12.Google Scholar
Kushibiki, S 2011. Tumor necrosis factor-α-induced inflammatory responses in cattle. Animal Science Journal 82, 504–511.Google Scholar
Livak, KJ and Schmittgen, TD 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25, 402–408.Google Scholar
Medzhitov, R 2007. Recognition of microorganisms and activation of the immune response. Nature 449, 819–826.Google Scholar
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, 998–1003.Google Scholar
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, 49–57.Google Scholar
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, 1340–1349.Google Scholar
Mukaida, N 2003. Pathophysiological roles of interleukin-8/CXCL8 in pulmonary diseases. American Journal of Physiology. Lung Cellular and Molecular Physiology 284, L566–L577.Google Scholar
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), ii17–ii21.Google Scholar
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, 399–409.Google Scholar
Pasare, C and Medzhitov, R 2004. Toll-like receptors: linking innate and adaptive immunity. Microbes and Infection 6, 1382–1387.Google Scholar
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, 1–23.Google Scholar
Rainard, P and Riollet, C 2006. Innate immunity of the bovine mammary gland. Veterinary Research 37, 369–400.Google Scholar
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, 6483–6502.Google Scholar
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, 2479–2486.Google Scholar
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, 3626–3635.Google Scholar
Sordillo, LM and Streicher, KL 2002. Mammary gland immunity and mastitis susceptibility. Journal of Mammary Gland Biology and Neoplasia 7, 135–146.Google Scholar
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, 146–152.Google Scholar
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, 2051–2056.Google Scholar
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, research0034.1–research0034.12.Google Scholar
Wellnitz, O and Bruckmaier, RM 2012. The innate immune response of the bovine mammary gland to bacterial infection. The Veterinary Journal 192, 148–152.Google Scholar
Yu, L, Wang, L and Chen, S 2010. Endogenous toll-like receptor ligands and their biological significance. Journal of Cellular and Molecular Medicine 14, 2592–2603.Google Scholar
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, 205–224.Google Scholar
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, 759–764.Google Scholar