Abstract

Brucellosis caused by Brucella organisms is a major zoonosis globally. It causes heavy losses through abortions, delayed conception and infertility in animals. Antibiotic therapy is ineffective. Once infected, the animal remains carrier and sheds bacteria in milk, semen and uterine discharges for long period. We have successfully used a lytic brucella phage for therapy of Brucellosis in adult cattle. We also targeted the phage employing the live attenuated Brucella abortus strain 19 organisms to kill the virulent Brucella residing intracellularly in phagocytes in the body. The effect of both the therapies was monitored non-invasively employing Brucella RNA in blood plasma as a Brucella - specific biomarker. A single dose of the phage alone or the therapeutic vaccine (phage pulsed S-19) could substantially reduce and finally eliminate live Brucella in the body within 3 months as evident from diminished and ultimately non – detectable RNA characteristic of Brucella abortus (223 bp amplicon) in plasma by RT-PCR. Thus, phage has a potential to cure Brucellosis and abolish carrier state in cattle and RNA can serve as a specific biomarker of live Brucella for monitoring and assessment of the efficacy of the therapy.

Keywords: Therapeutic Vaccine; Brucellosis; Brucella; Brucella Phage; Phage Therapy; Phage Targeting

References

1. Kollannur JD., et al. “Epidemiology and economics of brucellosis in animals and its zoonotic significance”. Proceedings of XIII International Congress in Animal Hygiene”. International Society for Animal Hygiene (2007): 466-468.
2. Jain U., et al. “Outbreak of brucellosis in buffaloes aborted in a village Mahuan, District Mainpuri, UP, India-A case report”. Veterinary World 6.1 (2013): 51-52.
3. Pandeya YR., et al. “Seroprevalence of brucellosis in different animal species of Kailali district Nepal”. International Journal of Infection and Microbiology 2 (2013): 22-25.
4. Godfroid J., et al. “Brucellosis in terrestrial wildlife”. Science Technology Review OIE 32.1 (2013): 27-42.
5. Prajapati A., et al. “Therapeutic efficacy of Brucellaphage against Brucella abortus in mice model”. Veterinary World 7.1 (2014): 34-37.
6. Haq IU., et al. “Bacteriophages and their implications on future biotechnology: A review”. Virology Journal 9 (2012): 9.
7. Mohan A and Saxena HM. “Effects of Bacteriophage Therapy on Host Immune Responses in Brucellosis Affected Cattle”. Acta Scientific Microbiology 3.6 (2020): 146-151.
8. Gupta V and Saxena HM. “Isolation and characterization of BpL1, a broad acting lytic bacteriophage against Brucella”. International Journal of Current Microbiology and Applied Sciences 6.11 (2017a): 2486-2496.
9. McDuff CR., et al. “Phenotypic alterations in the colonial morphology of Brucella abortus due to a bacteriophage carrier state”. Journal of Bacteriology 83 (1962): 860-866.
10. Rawat M and Verma R. “Isolation and characterization, preservation and therapeutic use of bacteriophage against Staphylococcus aureus associated with ruminant mastitis progress report”. Department of Biotechnology. Delhi. (BT/PR4 194/AAQ/ 158/ 2003).
11. Indian Pharmacopoeia. Government of India, Ministry of Health and Family Welfare. Indian Pharmacopoeia Commission, Ghaziabad (2010).
12. Morgan WJ., et al. Brucellosis diagnosis: Standard Laboratory Techniques Report Series no. 1, Weybridge, England (1978).
13. Baily GG., et al. “Detection of Brucella melitensis and Brucella abortus by DNA amplification”. Journal of Tropical Medicine and Hygiene 95 (1992): 271-275.
14. Gupta V and Saxena HM. “A new bacteriophage-based luminescence assay for diagnosis of Brucellosis”. Indian Journal of Experimental Biology 55 (2017b): 296-302.
15. Sheridan GE., et al. “Detection of mRNA by reverse transcription- PCR as an indicator of viability in Escherichia coli cells”. Applied and Environmental Microbiology 64 (1998): 1313-1318.
16. Deere D., et al. “Survival of cells and DNA of Aeromonas salmonicida released into aquatic microcosms”. Journal of Applied Bacteriology 81 (1996): 309-318. 
17. Hellyer TJ., et al. “Quantitative analysis of mRNA as a marker for viability of Mycobacterium tuberculosis”. Journal of Clinical Microbiology 37 (1999): 290-295.
18. Masters CI., et al. “Effect of stress treatments on the detection of Listeria monocytogenes and enterotoxigenic Escherichia coli by the polymerase chain reaction”. Journal of Applied Bacteriology 77 (1994): 73-79.
19. Keer JT and Birch L. “Molecular methods for the assessment of bacterial viability”. Journal of Microbiological Methods 53 (2003): 175-183.
20. Ling Yu W and Nielsen K. “Review of detection of Brucella spp. by Polymerase chain reaction”. Croatian Medical Journal 51 (2010): 306-313.
21. Mukherjee F., et al. “Multiple genus- specific markers in PCR assays improve the specificity and sensitivity of diagnosis of brucellosis in field animals”. Journal of Medical Microbiology 56 (1978): 1309-1316.
22. Elfaki MG., et al. “Detection of Brucella DNA in sera from patients with brucellosis by polymerase chain reaction”. Diagnostic Microbiology and Infectious Disease 53 (2005): 1-7.
23. Al-Ajlan HH., et al. “Comparison of different PCR methods for detection of Brucella species in human blood samples”. Polish Journal of Microbiology 60.1 (2011): 27-33.
24. Long GY., et al. “Immunization of rabbits against Pasteurellosis”. Chinese Journal of Veterinary Science and Technology's 2 (1986): 3-7.

Citation

Citation: Anju Mohan and Hari Mohan Saxena. “Non - Invasive Monitoring and Assessment of Phage Therapy and Phage Targeting in Bovine Brucellosis Employing a Brucella - Specific Biomarker". Acta Scientific Microbiology 3.9 (2020): 59-68.

Copyright

Copyright: © 2020 Anju Mohan and Hari Mohan Saxena. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.