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R esearch
(Published online:
16-03-2015)
14.
Neutrophil dynamics in the blood and milk of
crossbred cows naturally infected with
Staphylococcus aureus -
Dilip K. Swain,
Mohar Singh Kushwah, Mandheer Kaur and Ajay K. Dang
Veterinary World, 8(3): 336-345
doi:
10.14202/vetworld.2015.336-345
Dilip
K. Swain:
Division of Dairy Cattle Physiology, Lactation and
Immunophysiology Laboratory, National Dairy Research Institute,
Karnal - 132 001, Haryana, India;
dilip_swain@yahoo.com
Mohar
Singh Kushwah:
Division of Dairy Cattle Physiology, Lactation and
Immunophysiology Laboratory, National Dairy Research Institute,
Karnal - 132 001, Haryana, India;
drmoharvet@gmail.com
Mandheer Kaur:
Division of Dairy Cattle Physiology, Lactation and
Immunophysiology Laboratory, National Dairy Research Institute,
Karnal - 132 001, Haryana, India;
mandheerbiotech@yahoo.com
Ajay K.
Dang: Division of Dairy Cattle Physiology, Lactation and
Immunophysiology Laboratory, National Dairy Research Institute,
Karnal - 132 001, Haryana, India;
rajadang@gmail.com
Received:
02-11-2014, Revised: 31-01-2015, Accepted: 09-02-2015, Published
online: 16-03-2015
Corresponding author:
Parveen Kumar, e-mail: vetsparveen@gmail.com
Citation:
Swain DK, Kushwah MS,
Kaur M, Dang AK (2015) Neutrophil dynamics in the blood and milk
of crossbred cows naturally infected with Staphylococcus aureus,
Veterinary World 8(3):336-345.
Abstract
Aim:
The present study was designed
to evaluate the neutrophil dynamics in terms of the functional
competence during subclinical mastitis (SCM) and clinical mastitis
(CM).
Materials and Methods: A total
of 146 Karan fries cows were screened and were divided into three
groups as control (n=12), SCM, n=12 and CM, n=12 groups on the
basis of California mastitis test scoring, bacteriological
evaluation, gross and morphological changes in milk and by
counting milk somatic cell count (SCC). Both blood and milk
polymorphonuclear neutrophils (PMNs) were isolated in the study.
Phagocytic activity (PA) was studied by spectrophotometrically;
neutrophil extracelluar traps (NETs) were studied by scanning
electron microscopy (SEM); CD44 was quantified by flow cytometry
and apoptosis was studied by fluorescent microscopy.
Results: Significantly
(p<0.05) higher SCC, PA was found in milk of CM cows as compared
to SCM and control cows. Significantly lower (p<0.05) apoptosis
was observed in PMNs isolated from both blood and milk of CM group
of cows when compared to control and SCM group. The milk
neutrophils of CM group of cows formed NETs as evidenced from the
SEM images. Surface expression of CD44 revealed a significantly
(p<0.05) lower expression in milk neutrophils of CM group of cows
when compared to SCM and control group of cows.
Conclusion: The study
indicated a positive correlation between delayed neutrophil
apoptosis, persistent staying of neutrophils at the site of
infection along with formation of NETs as the strategies to fight
against the pathogens in the udder during
Staphylococcal mastitis.
The study forms a strong base
for future molecular research in terms of neutrophil recruitment
and neutrophil removal from the site of infection.
Keywords:
apoptosis, CD44, mastitis, neutrophil, neutrophil extracelluar
traps, Staphylococcus aureus.
References
1. Paape, M.J., Bannerman, D.D., Zhao, X. and Lee, J.W. (2003)
The bovine neutrophil: Structure and function in blood and
milk. Vet. Res., 34(5): 597-627.
http://dx.doi.org/10.1051/vetres:2003024
PMid:14556697 |
|
|
2. Borregaard, N. (2010) Neutrophils, from marrow to microbes.
Immunity, 33(5): 657-670.
http://dx.doi.org/10.1016/j.immuni.2010.11.011
PMid:21094463 |
|
|
3. Michael, A.R. and Stephen, C.N. (2011) Mastitis and its
impact on structure and function in the ruminant mammary
gland. J Mammary Gland Biol. Neoplasia., 16(4), 275-289.
http://dx.doi.org/10.1007/s10911-011-9231-3
PMid:21968535 |
|
|
4. Rainard, P. and Riollet, C. (2006) Innate immunity of the
bovine mammary gland. Vet. Res., 37(3): 369-400.
http://dx.doi.org/10.1051/vetres:2006007
PMid:16611554 |
|
|
5. Petzl, W., Zerbe, H.J., Guntler, J., Yang, W. and Seyfert,
H.M. (2008) Escherichia coli, but not Staphylococcus aureus
triggers an early increased expression of factors contributing
to the innate defense in the udder of the cow. Vet. Res.,
39(2): 18-26.
http://dx.doi.org/10.1051/vetres:2007057
PMid:18258172 |
|
|
6. Bratton, D.L. and Henson, P.M. (2011) Neutrophil clearance:
When the party is over, clean-up begins. Trends Immunol.,
32(8): 350-357.
http://dx.doi.org/10.1016/j.it.2011.04.009
PMid:21782511 PMCid:PMC3151332 |
|
|
7. Sadik, C.D., Kim, N.D. and Luster, A.D. (2011) Neutrophils
cascading their way to inflammation. Trends Immunol., 32(10):
452-460.
http://dx.doi.org/10.1016/j.it.2011.06.008
PMid:21839682 PMCid:PMC3470857 |
|
|
8. Kennedy, A.D. and DeLeo, F.R. (2009) Neutrophil apoptosis
and the resolution of infection. Immunol. Res., 43: 25-61.
http://dx.doi.org/10.1007/s12026-008-8049-6
PMid:19066741 |
|
|
9. Rainard, P., Fromageau, A., Cunha, P. and Gilbert, F.B.
(2008) Staphylococcus aureus lipoteichoic acid triggers
inflammation in the lactating bovine mammary gland. Vet. Res.,
39(5): 52-58.
http://dx.doi.org/10.1051/vetres:2008034
PMid:18593548 |
|
|
10. Brenaut, P., Lefèvre, L., Rau, A., Laloë, D., Pisoni, G.,
Moroni, P., Bevilacqua, C., and Martin, P. (2014) Contribution
of mammary epithelial cells to the immune response during
early stages of a bacterial infection to Staphylococcus
aureus. Vet. Res., 45: 16.
http://dx.doi.org/10.1186/1297-9716-45-16
PMid:24521038 PMCid:PMC3937043 |
|
|
11. Leitner, G., Lubashevsky, E., Glickman, A., Winkler, M.
and Trainin, Z. (2003) Development of a Staphylococcus aureus
vaccine against mastitis in dairy cows- challenge trials. Vet.
Immunol. Immunopathol., 93(1-2): 31-38.
http://dx.doi.org/10.1016/S0165-2427(03)00051-5 |
|
|
12. Swain, D.K., Kushwah, M.S., Kaur, M., Patbandha, T.K.,
Mohanty, A.K. and Dang, A.K. (2014) Formation of NET,
phagocytic activity, surface architecture, apoptosis and
expression of toll like receptors 2 and 4 (TLR2 and TLR4) in
neutrophils of mastitic cows. Vet. Res. Commun., 38: 209-219.
http://dx.doi.org/10.1007/s11259-014-9606-1
PMid:24915786 |
|
|
13. Mehrzad, J., Duchateau, L. and Burvenich, C. (2004)
Viability of milk neutrophils and severity of bovine coliform
mastitis. J. Dairy Sci., 87(12): 4150-4162.
http://dx.doi.org/10.3168/jds.S0022-0302(04)73558-4 |
|
|
14. Dang, A.K., Prasad, S., De, K., Mukherjee, J., Sandeep,
I.V.R., Mutoni, G., Pathan, M.M., Jamwal, M., Kapila, S.,
Kapila, R., Kaur, H., Dixit, S., Mohanty, A.K. and Prakash,
B.S. (2013) Effect of supplementation of vitamin E, copper and
zinc on the in vitro phagocytic activity and lymphocyte
proliferation index of peripartum Sahiwal (Bos indicus) cows.
J. Anim. Physiol. Anim. Nutr., 97(2), 315-321.
http://dx.doi.org/10.1111/j.1439-0396.2011.01272.x
PMid:22289079 |
|
|
15. Pyorala, S. (2003) Indicators of inflammation in the
diagnosis of mastitis. Vet. Res., 34(5): 565-578.
http://dx.doi.org/10.1051/vetres:2003026
PMid:14556695 |
|
|
16. Aitken, S.L., Corl, C.M. and Sordillo, L.M. (2011)
Immunopathology of mastitis: Insights into disease recognition
and resolution. J. Mammary Gland Biol. Neoplasia., 16(4):
291-304.
http://dx.doi.org/10.1007/s10911-011-9230-4
PMid:21938490 |
|
|
17. Sharma, N., Singh, N.K. and Bhadwal, M.S. (2011)
Relationship of somatic cell count and matitis; an overview.
Asian Aust. J. Anim. Sci., 24(3): 429-438.
http://dx.doi.org/10.5713/ajas.2011.10233 |
|
|
18. Kebir, D.E.I. and Filep, J.G. (2013) Targeting neutrophil
apoptosis for enhancing the resolution of inflammation. Cell,
2(2): 330-348.
http://dx.doi.org/10.3390/cells2020330
PMid:24709704 PMCid:PMC3972676 |
|
|
19. Boutet, P., Boulanger, D., Gillet, L. and Bureau, F.L.
(2004) Delayed neutrophil apoptosis in bovine subclinical
mastitis. J. Dairy Sci., 87(12): 4104-4114.
http://dx.doi.org/10.3168/jds.S0022-0302(04)73553-5 |
|
|
20. Lotz, S., Aga, S., Wilde, I., Zandbergen, G., Hartung, T.,
Solbach, W. and Laskay, T. (2004) Highly purified lipoteichoic
acid activates neutrophil granulocytes and delays their
spontaneous apoptosis via CD14 and TLR2. J. Leukocyte Biol.
75(3): 467-477.
http://dx.doi.org/10.1189/jlb.0803360
PMid:14673018 |
|
|
|
21. Sladek, Z., Rysanek, D. and Faldyna, M. (2005a) Effect of
Staphylococcus aureus and Streptococcus uberis on apoptosis of
bovine mammary gland tissue pool neutrophils in vitro. Vet.
Med., 50: 11-23. |
|
|
22. Sladek, Z., Rysanek, D., Ryznarova, H. and Faldyna, M.
(2005b) Neutrophil apoptosis during experimentally induced
Staphylococcus aureus mastitis. Vet. Res., 36(4): 629-643.
http://dx.doi.org/10.1051/vetres:2005023
PMid:15955286 |
|
|
23. Yamamoto, A., Taniuchi, S., Tsuji, S., Mausi, M. and
Kobayashi, Y. (2002) Role of reactive oxygen species in
neutrophil apoptosis following ingestion of heat-killed
Staphylococcus aureus. Clin. Exp. Immunol., 129(3): 479-484.
http://dx.doi.org/10.1046/j.1365-2249.2002.01930.x
PMid:12197889 PMCid:PMC1906464 |
|
|
24. Nathan, C. (2006) Neutrophils and immunity: Challenges and
opportunities. Nat. Rev. Immunol., 6(3): 173-182.
http://dx.doi.org/10.1038/nri1785
PMid:16498448 |
|
|
25. Brinkmann, V., Laube, B, Abu Abed, U., Goosmann, C. and
Zychlinsky, A. (2010) Neutrophil extracellular traps: how to
generate and visualize them. J. Vis. Exp., 36: 1724.
PMid:20182410 |
|
|
26. Papayannopoulos, V. Zychlinsky, A. (2009) NETs: A new
strategy for using old weapons. Trends Immunol., 30(11):
513-521.
http://dx.doi.org/10.1016/j.it.2009.07.011
PMid:19699684 |
|
