Isolation Pseudomonas aeruginosa bacteria and genes integron class I of subclinical mastitis in dairy cows in Tehran

Document Type : Full Research Paper

Authors

1 Department of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.

2 Professor, Department of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.

3 Assistant Professor, Department of Microbiology, Saveh Branch, Islamic Azad University, Saveh, Iran.

4 Professor, Department of Microbiology, Pasteur Institute of Iran, Tehran, Iran.

Abstract

Pseudomonas aeruginosa is an important opportunistic pathogen that causes subclinical mastitis resistant to treatment, in lactating dairy cows. The Purpose of this study, were to identify class I integrons and determining antibiotic susceptibility of Pseudomonas aeruginosa isolated from  mastitis. In this cross-sectional study, 150 samples of raw milk were collected from different farms, transported to the laboratory and cultured on specific media. Antimicrobial susceptibility test conducted  by disk diffusion method and MIC (Minimum Inhibitory Concentration) determination test performed by E-test in accordance with CLSI guidelines, in various groups. To identify Class I integron gene PCR test were used. Of total number 150 milk samples, 51 samples (34%) were contaminated with Pseudomonas aeruginosa. Antibiotic test results showed that the highest rate of resistance were observed to Ampicillin, Tetracycline and Kanamycin at the rate of 100%, 94.2% and 94.2%, respectively. Most multiple resistances were observed against Kanamycin, Ampicillin and Tetracycline antibiotics. MIC results showed that isolated bacteria were sensitive to Gentamicin in concentration 0.75-2 μg, Ciprofloxacin at 0.094-0.38 μg and Amikacin at 0.05-4 μg. Results molecular tset showed, one isolate (1.96%) were carried Integrons gene (int-1). Since most integrons genes coding enzymes that inactive antibiotics, presence of Integrons genes in samples of raw milk, treatment infections caused by Pseudomonas aeruginosa wich carring these genes, because of the lack of new antibiotics that have a broad therapeutic range, would creat the major problems. 

Keywords


1- Aertsen, A., K. Vanoirbeek, et al. (2004). Heat shock protein-mediated resistance to high hydrostatic pressure in Escherichia coli; Appl Environ Microbiol; 70(5): 2660-2666.
2- Aslani MM. Hashemipour M. Nikbin NS, S. F., Eidi A, Sharafi Z. (2009). Molecular detection and identification of virulence factors of Pseudomonas aeruginosa strains isolated from wound infections and burns; J Lorestan Uni Med Sci; 11(2): 23-29.
3- Brooks, G. F., J. S. Butel, et al. (2005). Jawetz, Melnick and Adelberg's medical microbiology; New York, NY, McGraw-Hill.
4- Champagne, C. P., R. R. Laing, et al. (1994). Psychrotrophs in dairy products: their effects and their control; Crit Rev Food Sci Nutr; 34(1): 1-30.
5- Cockerill, F. R., Clinical, et al. (2012). Performance standards for antimicrobial susceptibility testing: twenty-second informational supplement; provides updated tables for M02-A11 and M07-A9, National Committee for Clinical Laboratory Standards.
6- Daly, M., E. Power, et al. (1999). Molecular analysis of Pseudomonas aeruginosa: epidemiological investigation of mastitis outbreaks in Irish dairy herds; Appl Environ Microbiol; 65(6): 2723-2729.
7- Dubois, V., C. Arpin, et al. (2002). Clinical strain of Pseudomonas aeruginosa carrying a blaTEM-21 gene located on a chromosomal interrupted TnA type transposon; Antimicrob Agents Chemother; 46(11): 3624-3626.
8- Fluit, A. and F. Schmitz (1999). Class 1 integrons, gene cassettes, mobility, and epidemiology; Eur J Clin Microbiol Infect Dis; 18(11): 761-770.
9- Fluit, A. and F. J. Schmitz (2004). Resistance integrons and super‐integrons; Clin Microbiol Infect; 10(4): 272-288.
10- Fonseca, É. L., V. V. Vieira, et al. (2005). Class 1 integrons in Pseudomonas aeruginosa isolates from clinical settings in Amazon region, Brazil; FEMS Immunol Med Microbiol; 44(3): 303-309.
11- Ghaboli Mehrbani R, M. S., Khakpour M (2012). Assess the antibiotic susceptibility of bacteria causing mastitis in dairy cows dry period in Tabriz; J Vet Res Lab; 4(1): 140.
12- Gu, B., M. Tong, et al. (2007). Prevalence and characterization of class I integrons among Pseudomonas aeruginosa and Acinetobacter baumannii isolates from patients in Nanjing, China; J Clin Microbiol; 45(1): 241-243.
13- Jiang, Q., Y. Yang, et al. (2015). Microbial diversity analysis of subclinical mastitis in dairy cattle in Northeast China; African J Microbiol Res; 9(10): 687-694.
14- Khan, A. A. and C. E. Cerniglia (1994). Detection of Pseudomonas aeruginosa from clinical and environmental samples by amplification of the exotoxin A gene using PCR; Appl Environ Microbiol; 60(10): 3: 739-745.
15- Kirk, J. and R. Mellenberger (1987). Pseudomonas-infected dairy cows; Extension bulletin E-Cooperative Extension Service.
16- Kong, K.-F., S. R. Jayawardena, et al. (2005). Pseudomonas aeruginosa AmpR is a global transcriptional factor that regulates expression of AmpC and PoxB β-lactamases, proteases, quorum sensing, and other virulence factors; Antimicrob Agents Chemother: 49(11): 4567-4575.
17- Nazik, H., B. Ongen, et al. (2007). Genotype and antibiotic susceptibility patterns of Pseudomonas aeruginosa and Stenotrophomonas maltophilia isolated from cystic fibrosis patients; Jpn J Infect Dis; 60(2/3): 82.
18- Resmini, P., Pellegrino, L., & Battelli, G. (1990). Accurate quantification of furosine in milk and dairy products by a direct HPLC method; Ital J Food Sci; 2(3), 173-183.
19- Rowe-Magnus, D. A. and D. Mazel. (2002). The role of integrons in antibiotic resistance gene capture; Inter J Medl Microbiol; 292(2): 115-125.
20- Shem, M., J. Malole, et al. (2001). Incidence and causes of sub-clinical mastitis in dairy cows on smallholder and large scale farms in tropical areas of Tanzania; Asian Australas J Anim Sci; 14(3): 372-377.
21- Shojapour M, V. M., Shariati L, Karimi A, Zamanzad B. (2011). Determination of antibiotic resistance and check-lactamase producing Pseudomonas aeruginosa strains isolated from clinical specimens Kashani and Hajar hospitals in 1387; South Med J; 14(2): 99-94.
22- Teale, C., P. Martin, J. Rogers, and Mars, A. (2004). VLA Antimicrobial Sensitivity Report.2003; HMSO: 23-24.
23- Zadoks, R. N., J. R. Middleton, et al. (2011). Molecular epidemiology of mastitis pathogens of dairy cattle and comparative relevance to humans; J Mamm Gland Biol Neoplasia; 16(4): 357-372.
24- Zeighami H, H. F., Hajiahmadi F (2013). The role of integrons in the development of antibiotic resistance; Lab Diagn; 92: 61-71.