Immunomodulation of IL-1, IL-6 and IL-8 cytokines by Prosopis juliflora alkaloids during bovine sub-clinical mastitis
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The current work is focused on establishing therapeutic protocol using unconventional drugs of herbal origin and studying their mechanism of action at molecular level in the treatment of bovine sub-clinical mastitis. It explores the potential of different cytokines which can be used for diagnosis, prognosis and monitoring of bovine sub-clinical mastitis. Prosopis juliflora alkaloids was administered intramammarily in 24 sub-clinically affected quarters once a day for 5 consecutive days at the rate of 10 ml of 1% formulation. In 18 disease control quarters, sterile normal saline was infused. The bacterial cultural examination, somatic cell count (SCC) and cytokines (IL-1β, IL-6, IL-8, IL-12, GM-CSF, IFN-γ, TNF-α) expression by real-time PCR were evaluated on day 7, 14, 21 and 28 post-last treatment from milk samples. Around 75.0% of treatment group quarters showed significant (p < 0.05) reduction in SCC on day 28 post-last treatment, whereas 94.4% control group quarters did not show any significant decline in SCC. 58.3% of treated quarters showed both bacteriological cure as well as significant (p < 0.05) reduction in SCC on day 28 post-last treatment. While, among control group quarters, 83.3% quarters not only remained bacteriological positive, they also did not show any significant decline in SCC. The in vitro antimicrobial activity of alkaloids of P. juliflora was evaluated. Lower concentrations of alkaloids (0.25% and 0.50%) dissolved in normal saline showed zone of inhibition against 12 out of 15 isolates, however higher concentration (1, 1.5, 2, 2.5 and 5%) showed zone of inhibition against all 15 bacterial isolates. The gene expression level of IL-1β, IL-8 and IFN-γ cytokines exhibited significant difference between healthy and sub-clinically affected quarters highlighting the potential of these cytokines in the diagnosis of bovine sub-clinical mastitis. Down-regulation of IL-1, IL-6, IL-8 and IFN-γ cytokines in treated quarters can be explored for making the prognosis and monitoring post-treatment disease progression of bovine sub-clinical mastitis. The P. juliflora alkaloid demonstrated strong in vitro and in vivo antibacterial activity, along with causing immunomodulation by enhancing post-treatment gene expression of IL-1, IL-6 and IL-8 cytokines. Therefore, P. juliflora alkaloids hold a strong claim as an effective alternative herbal therapy in bovine sub-clinical mastitis.
KeywordsAlkaloids Bovine sub-clinical mastitis Cytokines Prosopis juliflora Real-time PCR Somatic cell count Treatment
The authors are thankful to Government of India’s Rashtriya Krishi Vikas Yojana (RKVY) Stream-I Project (B.H. 18442) for providing financial support. KS is appreciative of Dr. Vaibhav Bhatt, Dr. Pradeep Kumar Sharma, Dr. Maulik Upadhyay and Dr. Kamlesh Sadaria for the invaluable interactions. KS expresses his gratitude towards the milkers of Livestock Research Station of Anand Agricultural University for extending their support during the entire study.
This study was funded by Rashtriya Krishi Vikas Yojana (RKVY) (Stream-I), Government of India. Money earmarked for RKVY projects is given to Government of Gujarat, which is allocated to different universities by former. The funds were allocated by Anand Agricultural University under the Budget Head (B.H.) 18442.
Compliance with ethical standards
Based on the results of the acute dermal and oral toxicity test carried out on male Wistar rats (as per OECD New Draft Guideline 434) the use of P. juliflora alkaloids was approved by the Institution Animal Ethics Committee (IAEC 2007/VPT/26). All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.
Conflict of interest
Prof. Dr. Dev S. Nauriyal has received research grant from Rashtriya Krishi Vikas Yojana (RKVY) (Stream-I), Government of India, as Principal Investigator for project entitled “Use of Recent Molecular Techniques to Reduce Economic Losses Incurred due to Bovine Mastitis by Evolving Diagnostic, Therapeutic and Preventive Measures” under the B.H. 18442. Prof. Dr. C. G. Joshi is Co-PI in this project. The authors declare that they have no conflict of interest.
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