Resistant Pattern of Therapeutics Antimicrobial Challenged on Pseudomonas aeruginosa Bacterium Isolated from Marketed Raw Buffalo Milk

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N. M. Patel
R. Kumar
A. P. Suthar
D. N. Desai
I. H. Kalyani


The study was designed to isolate predominance contamination of Pseudomonas aeruginosa in marketed raw buffalo milk (n=122) samples, collected from private dairy farms from different places of south Gujarat, India. Pre-enrichment of 1 ml of each sample was done with inoculation in 9 ml tryptone soya broth and incubated at 37ºC for 24 hrs. A loopful of culture was taken from broth and streaked on selective Pseudomonas agar F plates and incubated at 37ºC for 24 hrs, after completion of incubation period, the colonies characteristics were studied and further confirmed by various biochemical tests and found 14 samples contaminated with P. aeruginosa, were further more biochemical testes are used and give positive results with IMViC, Motility test, catalase and sugar fermentation confirm at 37ºC for 24 hrs incubation. All biochemically conformed isolates were further subjected for molecular characterization and were also tested for antimicrobial susceptibility by using various antibiotics discs such as vancomycin, penicillin, tylosin, cefixime, chloramphenicol, kanamycin, ceftriaxone, cefixime, tetracycline, streptomycin, ciprofloxacin, enrofloxacin, oxytetracycline and gentamicin, which has shown multi drug resistant ranging from seven to nine antimicrobials and Multiple Antibiotic Resistance (MAR) index ranges from 0.50 to 0.64.The isolates of P. aeruginosa in the present study are extremely resistant to vancomycin, penicillin, tylosin, cefixime, chloramphenicol and maximum sensitive to ciprofloxacin and enrofloxacin followed by gentamicin. Further statistical analysis of antibiotics wise zone diameter interpretative standard (mm) reveled susceptibility phenotypes under significant of difference at P≤0.05 in one way ANOVA using Duncan’s multiple range test and found ciprofloxacin having maximum sensitivity among antibiotic tested and it could be considered as a drug of choice for controlling P. aeruginosa mediated animal and human infections in the studied regions for insuring food safety as well.

Pseudomonas aeruginosa, buffalo, milk, antimicrobial, resistant, therapeutics.

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How to Cite
Patel, N. M., Kumar, R., Suthar, A. P., Desai, D. N., & Kalyani, I. H. (2019). Resistant Pattern of Therapeutics Antimicrobial Challenged on Pseudomonas aeruginosa Bacterium Isolated from Marketed Raw Buffalo Milk. European Journal of Nutrition & Food Safety, 9(4), 398-407.
Original Research Article


Zhang SB, Zhai HC, Hu YS, Wang L, Yu GH, Huang SX, Cai JP. A rapid detection method for microbial spoilage of agro-products based on catalase activity. Food Control. 2014;42:220-224.

Bashir S, Awan MS, Khan SA, Suthar V, Khan MM, Khan MT. Microbiological quality evaluation of raw milk consumed in and around Rawalakot, Azad Jammu and Kashmir. International Journal of Microbiology Research. 2014;5:112-116.

Garedew L, Berhanu A, Mengesha D, Tesgay G. Dentification of gram-negative bacteria from critical control points of raw and pasteurized cow milk consumed at Gondar town and its suburbs, Ethiopia. BMC Public Health. 2012;12:950.

Kumar R, Singh SP, Kumar M, Kumar A. Determination of low molecular weight immunogenic Omp28 Protein and Gene of Salmonella typhimurium. South Asian Journal of Experimental Biology. 2018;8(5):172-177.

Singh P, Wani AA, Karim AA, Langowski AA. The use of carbon dioxide in the processing and packaging of milk and dairy products: A review. International Journal of Dairy Technology. 2012;65:161-177.

Olfa SB, Imène F, Rouaa L, Hamadi A, Mohamed AA. Study of proteolytic and lipolytic Activities of Pseudomonas spp. Isolated from pasteurized milk in Tunisia. Journal of Agricultural Science. 2013;5(7):46-50.

Aylin A, Evrim GA, Gokce PY. A modified selective medium containing Benzalkonium Chloride (BKC) for the isolation of Pseudomonas aeruginosa from raw milk. Food and Nutrition Sciences. 2012;3(7):947-950.

Wiedmann M, Weilmeier D, Dineen SS, Ralyea RM, Boor KJ. Molecular and phenotypic characterization of Pseudomonas spp. isolated from milk. Applied Environment Microbiology. 2000;66(5):2085-2095.

Osundiya OO, Oladele RO, Oduyebo OO. Multiple Antibiotic Resistance (MAR) indices of Pseudomonas and Klebsiella species isolates in Lagos University Teaching Hospital. African Journal of Clinical and Experimental Microbiology. 2013;14(3):164 -168.

Kumar R, Dutta S, Pal D. Microbiological quality assessment and detection of fecal Escherichia coli (Coliform) in foods of animal origin sold in the open markets of Kolkata city in India. Indian Journal of Comparative Microbiology Immunology and Infectious. Diseases. 2017;38(1):22-25.

Arakawa K, Kawai Y, Iioka H, Tanioka M, Nishimura J, Kitazawa H, Tsurumi K, Saito T. Microbial community analysis of food-spoilage bacteria in commercial custard creams using culture- dependent and independent methods. Journal of Dairy Science. 2008;91(8):2938-2946.

Kumar R, Singh SP. Pulse field gel electrophoresis-profiles of salmonella enteric serovar typhimurium isolates obtained from chickens sources. Indian Journal of Comparative Microbiology Immunology and Infectious, Diseases. 2014;35(20): 64-66.

Vos P, Garrity G, Jones D, Krieg NR, Ludwig W, Rainey FA, Schleifer KH, Whitman WB. [1984 (Williams & Wilkins)]. George M. Garrity (ed.). the firmicutes. Bergey's manual of systematic bacteriology. 3 (2nd ed.). New York: Springer. 2009;1450.
ISBN 978-0-387-95041-9. British Library no. GBA561951.

CLSI. Performance standards for antimicrobial susceptibility testing., 27th ed. CLSI supplement M100. Clinical and Laboratory Standards Institute, Wayne, PA 19087 USA; 2017.

Bauer AW, Kirby WM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology. 1966;45(4):4593-4596.

Piyush Tripathy, Gopa Banerjee, Shivani Saxena, Mahendra Kumar Gupta, Ramteke PW. Antibiotic resistance pattern of Pseudomonas aeruginosa isolated from patients of lower respiratory tract infection. African Journal of Microbiology Research. 2011;5(9):2955-2959.

Spilker T, Coenye T, Vandamme P, Li Puma JJ. PCR-based assay for differentiation of Pseudomonas aeruginosa from other Pseudomonas species recovered from cystic fibrosis patients. Journal of Clinical Microbiology. 2004;42(5):2074-2079.

Nawaz MA, Bhattarai RR. Isolation and identification of Yersinia and Pseudomonas spp. from Australian milk and salad using 16S rDNA. Peer J. Pre Prints. 2015;3:e820v1.

El-Zubeir IEM, El-Owni OAO. Antimicrobial resistance of bacteria associated with raw milk contaminated by chemical preservatives. World Journal of Dairy & Food Sciences. 2009;4 (1):65-69.

Jyoti Y, Saurav P, Jyotsna KP, Yashab K, Ajay K, Singh Florin M, Harison M. Comparative evaluation of pathogenic bacterial incidence in raw and pasteurized milk. International Journal of Engineering Science Invention. 2014;3(5):11-20.

Jombo GTA, Jonah P, Ayeni JA. Multidrug resistant Pseudomonas aeruginosa in contemporary medical practice: Findings from urinary isolates at a Nigerian University Teaching Hospital. Nigerian Journal of Physiological Sciences. 2008;23(1-2):105-109.

Hajira Bilal, Fariha Hasan, Samina Bilal. Susceptibility pattern of Pseudomonas aeruginosa against various antibiotics along with computational analysis. International Journal of Sciences: Basic and Applied Research. 2015;24(4):23-45.

Gedamu Shewatatek, Gizachew Tilahun, Molalegne Bitew, Terefe Gelibo. Drug sensitivity of Pseudomonas aeruginosa from wound infections in Jimma University Specialized Hospital, Ethiopia. Online Journal of Medicine and Medicinal Research. 2014;3(2):13-18.

Garba I, Lusa H, Bawa E, Tijjani MB, Aliyu MS, Zango UU, Raji MIO. Antibiotic susceptibility pattern of Pseudomonas aeruginosa isolated from wounds in patients attending ahmadu bello university teaching hospital, Zaira, Nigeria. Nigerian Journal of Basic and Applied Science. 2012;20(1):32-34.

Tamil SS, Murugan S. Antimicrobial Susceptibility Patterns of Pseudomonas aeruginosa from diabetes patients with foot ulcers. International Journal of Microbiology. 2011;4.
Article ID 605195

Gad GF, Ramadan A, El-Domany, SZ, Hossam MA. Characterization of Pseudomonas aeruginosa isolated from clinical and environmental samples in Minia, Egypt: Prevalence, antibiogram and resistance mechanisms. Journal of Antimicrobial Chemotherapy. 2007;60: 1010-1017.

Okon KO, Aguwe PC, Oladosu W, Balogun Uba A. Antibiotic resistance patterns of Pseudomonas aeruginosa isolated from clinical specimens in a tertiary care hospital in North eastern Nigeria. Journal of Microbiology and Antimicrobials. 2009;1(2):19-26.

Al-Kabsi AM, Yusof M, Sekaran SH. Antimicrobial resistance pattern of clinical isolate of Pseudomonas aeruginosa in the University of Malaya Medical Center, Malaysia. African Journal of Microbiology. 2011;5:5266-5272.

Zaheer Ali, Nusrat M, Sehar ANNJ, Maryam S. Multi-drug resistant Pseudomonas aeruginosa: A threat of nosocomial infections in tertiary care hospitals. Journal of the Pakistan Medical Association. 2015;65(1):12-16.

Ahmed BM, Ahmed ZW, Rashad HG, Zaghlol LA, Galal R. Prevalence of Multi drug resistant Pseudomonas aeruginosa in patients with nosocomial infection at a University Hospital in Egypt, with special reference to typing methods. Journal of Virology and Microbiology. 2013;01-13.

Ahmad BM, Kudi MT. Chronic suppurative otitis media in Gombe, Nigeria. Nigerian Journal of Surgical Research. 2003;5(3):120-123.

Ferede D, Geyid A, Lulseged S, Melaku A. Drug susceptibility pattern of bacterial isolates from children with chronic suppurative otitis media. Ethiopian Journal of Health Development. 2001;15(2):89-96.

Ogundipeju OO, Nwobu RAU. Occurrence of Pseudomonas aeruginosa in post-operative wound infection. Pakisthan Journal of Medical. 2004;20:187-191.

Emmanuel EO, Etinosa Ol, Raphael M, Bright I, Clara LI, Anthony IO. Prevalence of Multiple Antibiotic Resistant (MAR) Pseudomonas species in the final effluents of three municipal waste water treatment facilities in south Africa. International Journal of Environmental Research and Public Health. 2012;9(6):2092-2107.

Mohammed GMO, Megahed AA, Nasr SS. Bacteriological and molecular detection of pseudomonas species from raw milk sold in Port-Said City markets. Egyptian Journal of Chemistry and Environmental Health. 2015;1(1):986-1002.