title: Flow cytometric analysis: Interdependence of healthy and infected udder quarters.
abstract: An important question about intramammary infections that is still debated in the literature is the independence or interdependence of the quarters of dairy cows. The present study sought to explore milk neutrophil function and the milk lymphocyte profile of uninfected quarters from uninfected and infected (one infected quarter per cow) udders to evaluate interdependence of the quarters. Thus, 32 (8 cows) and 18 (6 cows) uninfected quarters from uninfected and infected udders were used, respectively. Using flow cytometry, we evaluated the percentage of milk neutrophils and their expression of adhesion molecules L-selectin (CD62L), β2-integrin (CD11b), and an endothelial-selectin ligand (CD44); levels of intracellular reactive oxygen species (ROS); phagocytosis of Staphylococcus aureus by milk neutrophils; and neutrophil viability. Furthermore, we assessed the percentage of B-cell (CD21(+)) and T-lymphocyte subsets (CD3(+)/CD4(+)/CD8(-), CD3(+)/CD8(+)/CD4(-), CD3(+)/CD4(+)/CD25(-), CD3(+)/CD4(+)/CD25(+), and CD3(+)/CD4(-)/CD25(-)) using flow cytometry with monoclonal antibodies. The infected quarter did not affect somatic cell count or the percentage of neutrophils in the neighboring uninfected quarters. Furthermore, the infected quarter did not influence neutrophil viability, intracellular reactive oxygen species production, or phagocytosis of S. aureus by milk neutrophils. Conversely, the expression of adhesion molecules CD11b, CD62L, and CD44 by milk neutrophils differed between uninfected quarters from infected versus uninfected udders. The lymphocyte subsets did not differ between groups, except for a higher percentage of B cells in uninfected quarters from infected udders than in those from uninfected udders. Thus, our study strongly supports the hypothesis of interdependence of quarters based on the influence of infection on both the percentage of B cells and the expression of adhesion molecules by milk neutrophils in the neighboring uninfected quarters.
authors: Departamento de Clínica Médica, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87, Cidade Universitária, São Paulo 05508-270, Brazil. Electronic address: magblagitz@uol.com.br.
authors: M G
authors: MG
authors: Blagitz
authors: Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte 31270-010, Brazil.
authors: F N
authors: FN
authors: Souza
authors: Departamento de Clínica Médica, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87, Cidade Universitária, São Paulo 05508-270, Brazil.
authors: C F
authors: CF
authors: Batista
authors: Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte 31270-010, Brazil.
authors: S A
authors: SA
authors: Diniz
authors: Departamento de Clínica Médica, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87, Cidade Universitária, São Paulo 05508-270, Brazil.
authors: L F F
authors: LF
authors: Azevedo
authors: Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte 31270-010, Brazil.
authors: M X
authors: MX
authors: Silva
authors: Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte 31270-010, Brazil.
authors: J P A
authors: JP
authors: Haddad
authors: Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte 31270-010, Brazil.
authors: M B
authors: MB
authors: Heinemann
authors: Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte 31270-010, Brazil.
authors: M M O P
authors: MM
authors: Cerqueira
authors: Departamento de Clínica Médica, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87, Cidade Universitária, São Paulo 05508-270, Brazil.
authors: A M M P
authors: AM
authors: Della Libera
mesh_terms: D000818:Animals; D001402:B-Lymphocytes; D018821:CD18 Antigens; D002417:Cattle; D015815:Cell Adhesion Molecules / Q000032:analysis; D005260:Female; D005434:Flow Cytometry / Q000662:veterinary*; D019041:L-Selectin / Q000276:immunology; D007958:Leukocyte Count / Q000662:veterinary; D018655:Lymphocyte Count / Q000662:veterinary; D016131:Lymphocyte Subsets / Q000276:immunology; D008321:Mammary Glands, Animal / Q000276:immunology*; D008414:Mastitis, Bovine / Q000276:immunology* / Q000382:microbiology / Q000503:physiopathology; D008892:Milk / Q000166:cytology / Q000276:immunology; D009504:Neutrophils / Q000737:chemistry / Q000276:immunology; D010587:Phagocytosis / Q000276:immunology; D013203:Staphylococcal Infections / Q000662:veterinary; D013211:Staphylococcus aureus / Q000276:immunology; D013601:T-Lymphocytes
chemical_list: D018821:CD18 Antigens; D015815:Cell Adhesion Molecules; D019041:L-Selectin
keywords: dairy cow; lymphocyte; mastitis; neutrophil; somatic cell count