Open Access
Research (Published online: 04-07-2017)
4. Detection and characterization of pathogenic Pseudomonas aeruginosa from bovine subclinical mastitis in West Bengal, India
S. Banerjee, K. Batabyal, S. N. Joardar, D. P. Isore, S. Dey, I. Samanta, T. K. Samanta and S. Murmu
Veterinary World, 10(7): 738-742

S. Banerjee: Department of Veterinary Microbiology, Faculty of Veterinary and Animal Sciences, West Bengal University of Animal and Fishery Sciences, Kolkata, West Bengal, India.
K. Batabyal: Department of Veterinary Microbiology, Faculty of Veterinary and Animal Sciences, West Bengal University of Animal and Fishery Sciences, Kolkata, West Bengal, India.
S. N. Joardar: Department of Veterinary Microbiology, Faculty of Veterinary and Animal Sciences, West Bengal University of Animal and Fishery Sciences, Kolkata, West Bengal, India.
D. P. Isore: Department of Veterinary Microbiology, Faculty of Veterinary and Animal Sciences, West Bengal University of Animal and Fishery Sciences, Kolkata, West Bengal, India.
S. Dey: Department of Veterinary Microbiology, Faculty of Veterinary and Animal Sciences, West Bengal University of Animal and Fishery Sciences, Kolkata, West Bengal, India.
I. Samanta: Department of Veterinary Microbiology, Faculty of Veterinary and Animal Sciences, West Bengal University of Animal and Fishery Sciences, Kolkata, West Bengal, India.
T. K. Samanta: Avian Influenza Laboratory, Institute of Animal Health & Veterinary Biologicals (R&T), Government of West Bengal, Kolkata, West Bengal, India.
S. Murmu: Department of Veterinary Public Health, Faculty of Veterinary and Animal Sciences, West Bengal University of Animal and Fishery Sciences, Kolkata, West Bengal.

doi: 10.14202/vetworld.2017.738-742

Share this article on [Facebook] [LinkedIn]

Article history: Received: 25-01-2017, Accepted: 13-05-2017, Published online: 04-07-2017

Corresponding author: K. Batabyal

E-mail: drkb.micro@gmail.com

Citation: Banerjee S, Batabyal K, Joardar SN, Isore DP, Dey S, Samanta I, Samanta TK, Murmu S (2017) Detection and characterization of pathogenic Pseudomonas aeruginosa from bovine subclinical mastitis in West Bengal, India, Veterinary World, 10(7): 738-742.
Abstract

Aim: Subclinical mastitis in bovines is mainly responsible for the huge economic loss of the dairy farmers, of which Pseudomonas aeruginosa is one of the causative agents. The study was aimed at a screening of suspected milk samples from different cattle farms of West Bengal for detection and confirmation of P. aeruginosa strains followed by their characterization.

Materials and Methods: Around 422 milk samples were screened from different dairy farms primarily by on-spot bromothymol blue (BTB) test and then in the lab by somatic cell counts (SCC) to finally consider 352 samples for detection of P. aeruginosa. Selective isolation and confirmation of the isolates were done using selective media, viz., cetrimide and Pseudomonas agar followed by confirmation by fluorescent technique. Molecular characterization of the strains was done by polymerase chain reaction for the presence of toxA (enterotoxin A, 352 bp) and exoS (exoenzyme S, 504 bp) genes.

Results: Approximately, 371 (87.9%) samples were positive in on-spot BTB test among which 352 (94.8%) samples revealed high SCC values (more than 3 lakh cells/ml) showing infection when screened. Among these, 23 (6.5%) samples yielded typical Pseudomonas sp. isolates out of which only 19 (5.4%) isolates were confirmed to be P. aeruginosa which showed characteristic blue-green fluorescence due to the presence of pigment pyoverdin under ultraviolet light. Out of these 19 isolates, 11 isolates were positive for toxA, 6 isolates for exoS, and 2 for both these pathogenic genes.

Conclusion: Approximately, 5.4% cases of bovine subclinical mastitis infections in South Bengal were associated with P. aeruginosa which possess pathogenic genes such as toxA (63.2%) and exoS (36.8%).

Keywords: bovines, characterization, exoS, Pseudomonas aeruginosa, subclinical mastitis, toxA.

References

1. N.D.D.B. Statistics. (2013) Milk Production in India. IST; 2013. Available from: http://www.nddb.coop/English/Statistics/ Pages/ Milk-Production.aspx. Accessed on 22-10-2013.

2. Kumar, M., Goel, P., Sharma, A. and Kumar, A. (2009) In: Compendium of 27th ISVM International Summit and Convention at Chennai, Tamil Nadu, India. p4-6.

3. Dua, K. (2001) Incidence, etiology and estimated economic losses due to mastitis in Punjab and in India - An update. Indian Dairyman, 53(10): 41-48.

4. Patel, J.V., Bhingaradia, B.V., Patel, B.B., Patel, S.B., Patel, P.B. and Vahora, S.P. (2012) Study on prevalence of mastitis and antibiotic sensitivity of bacterial isolates recovered from crossbred cows of Anand district of Gujarat. Indian J. Dairy Sci., 65(6): 467-471.

5. Singh, R., Sharma, N., Soodan, J.S. and Sudhan, N.A. (2005) Etiology and sensitivity of bacterial isolates from sub-clinical mastitis in cattle from Jammu region. SKUAST J. Res., 4(2): 223-224.

6. Narayanan, S. (2013) Pseudomonas. In: McVey, D.S., Kennedy, M. and Chengappa, M.M., editors. Veterinary Microbiology. 3rd ed. John Wiley & Sons, Inc., New York. p148-150.

7. Chakrabarti, A. (2007) A Textbook of Preventive Veterinary Medicine. 4th ed. Kalyani Publishers, New Delhi. p477-480.

8. Sharma, N., Singh, N.K. and Bhadwal, M.S. (2011) Relationship of somatic cell count and mastitis: An overview. Asian Australas. J. Anim. Sci., 24: 429-438. [PubMed]

9. Smith, K.L. (1996) Standards for Somatic Cells in Milk: Physiological and Regulatory. Mastitis Newsletter, Newsletters of the IDF No. 144. p7-9.

10. Quinn, P.J., Markey, B.K., Leonard, F.C., Fitz Patrick, E.S., Fanning, S. and Hartigan, P.J. (2011) Veterinary Microbiology and Microbial Diseases. 2nd ed. Blackwell Publishing Ltd., Ames, IA. p287-290. [PubMed]

11. Carter, G.R. and Wise, D.J. (2004) Essentials of Veterinary Bacteriology and Mycology. 6th ed. The Iowa State Press, Iowa. p125-126.

12. Samanta, I. (2013) Pseudomonas and Burkholderia. In: Veterinary Bacteriology. 1st ed. Ch. 10. New India Publishing Agency, New Delhi. p209-223.

13. Lanotte, P., Watt, S., Mereghetti, L., Dartiguelongue, N., Rastegar-Lari, A., Goudeau, A. and Quentin, R. (2004) Genetic features of Pseudomonas aeruginosa isolates from cystic fibrosis patients compared with those of isolates from other origins. J. Med. Microbiol., 53: 73-81. [Crossref] [PubMed]

14. Brenner, D.J., McWhorter, A.C., Knutson, J.K. and Steigerwalt, A.G. (1982) Escherichia vulneris: A new species of Enterobacteriaceae associated with human wounds. J. Clin. Microbiol., 15: 1133-1140. [PubMed] [PMC]

15. Harini, H. and Sumathi, B.R. (2011) Screening of bovine milk samples for sub-clinical mastitis and antibiogram of bacterial isolates. Vet. World, 4(8): 358-359.

16. Marschke, R.J. and Kitchen, B.J. (1985) Detection of bovine mastitis by bromothymol blue pH indicator test. J. Dairy Sci., 68: 1263-1269. [Crossref]

17. Singh, R., Bansal, B.K., Uppal, S.K. and Malik, D.S. (2000) Diagnosis of sub-clinical mastitis: A comparative study of different tests. Indian J. Anim. Res., 34(2): 120-123.

18. Langer, A., Sharma, S., Sharma, N.K. and Nauriyal, D.S. (2014) Comparative efficacy of different mastitis markers for diagnosis of sub-clinical mastitis in cows. Int. J. Appl. Sci. Biotechnol., 2(2): 121-125. [Crossref]

19. Shitandi, A., Ogollah, H. and Nanua, J.N. (2005) Effect of sub-clinical mastitis on milk composition in the Kenyan smallholder dairy herds. Afr. Crop Sci. Conf. Proc., 7: 545-550.

20. Freitas, A.L. and Barth, A.L. (2004) Typing of Pseudomonas aeruginosa from hospitalized patients: A comparison of susceptibility and biochemical profiles with genotype. Braz. J. Med. Biol. Res., 37: 77-82. [Crossref] [PubMed]

21. Heleili, N., Ayachi, A., Melizi, M., Kassah, A.L. and Mamache, B. (2012) Prevalence of sub-clinical bovine mastitis and the in vitro sensitivity of bacterial isolates in Batna governorate, East of Algeria. J. Anim. Sci. Adv., 2(6): 576-582.

22. Viswakarma, P. (2008) Studies on Prevalence, Diagnosis, Therapy and Control of Mastitis in Buffaloes. M. V. Sc. Thesis. Indira Gandhi Agricultural University, Raipur, Chhattisgarh.

23. Badr, H.A.R., El-Nagdy, M., El-Sabagh, A. and El-Din, A.B. (2008) Pseudomonas aeruginosa exotoxin a as a virulence factor in burn wound infections. Egypt. J. Med. Microbiol., 17(1): 125-132.

24. Nikbin, V.S., Aslani, M.M., Sharafi, Z., Hashemipour, M., Shahcheraghi, F. and Ebrahimipour, G.H. (2012) Molecular identification and detection of virulence genes among Pseudomonas aeruginosa isolated from different infectious origins. Iran. J. Microbiol., 4(3): 118-123. [PubMed] [PMC]

25. Sharma, S., Kaur, R., Yadav, V., Harjai, K. and Joshi, K. (2004) Contribution of exotoxin A of Pseudomonas aeruginosa in acute & chronic renal infection. Jpn. J. Infect. Dis., 57: 119-120. [PubMed]