OJMM  Vol.4 No.2 , June 2014
Isolation and Identification of Multi-Drug Resistant Strains of Non-Lactose Fermenting Bacteria from Clinical Isolates
ABSTRACT
Purpose: We studied the drug resistance of different microbes from clinical isolates. The morphological characteristics of bacteria were observed through culture characteristics and by carrying out gram staining techniques while the biochemical characteristics of bacteria were carried out by biochemical test. Methods: A total of 324 samples were collected from suspected patients visiting different hospitals at district Peshawar. For morphological identification, samples of clinical isolates were analyzed by blood agar, MacConkey agar and Eosine Methylene Blue, identified by gram staining and characterized by different biochemical tests. Antibiotic Sensitivity test by Modified Kirby-Bauer Disc diffusion method was used to test the in-vitro susceptibility of the identified isolates to different antibiotics such as Ceftazidime, Ceftazidime, Ceftriaxone, Cefepime and Imipenem. Results: These resistant non-lactose fermenting gram negative bacteria were isolated from samples of pus/wound (33.30%, n = 108/324), blood (33.30%, n = 108/324), urine (23.30%, n = 75/324) and from ascetic/pleural fluids (10.20%, n = 33/324). The study revealed that the percentage of non-fermenting bacterial infection was higher in females (53%) as compared to males (47%) along with higher infection observed in the age group of 11 - 30 years. Pseudomonas aeroginosa showed high resistance against Cefepime (88.80%), followed by Cefoperazone (55.50%), Ceftazidime (48.10%), Ceftriaxone (33.30%). Imipenem was active with low resistance (7.40%). More resistance was seen in Morganella morganii against Imipenem (66.70%) followed by Cefoperazone (55.50%), Ceftriaxone (55.50%). Cefepime showed low resistance (11%). Multi-drug resistant Proteus mirabillis was highly resistance to Ceftriaxone (74.07%), followed by Cefepime (59.20%), Cefoperazone (44.40%) and low resistance for Imipenem (25.90%). Salmonella typhi demonstrated high resistance against Imipenem (74.07%), followed by Ceftriaxone (40.70%), Ceftazidime (37.03%). Cefepime showed low resistance (3.70%), hence it is more active against S. typhi. Conclusions: The different species of non-lactose fermenting gram negative bacteria have shown a different resistivity pattern in the present study. Therefore identification of non-lactose fermenting gram negative bacteria and looking after their resistivity/susceptibility pattern are important for suitable management of the infections caused by them.

Cite this paper
Akbar, M. , Zahid, M. , Ali, P. , Sthanadar, A. , Shah, M. , Sthanadar, I. , Kaleem, M. , Aslam, M. ,  , K. ,  , Z. , Hassan, S. , Jehan, N. and Khan, M. (2014) Isolation and Identification of Multi-Drug Resistant Strains of Non-Lactose Fermenting Bacteria from Clinical Isolates. Open Journal of Medical Microbiology, 4, 115-123. doi: 10.4236/ojmm.2014.42013.
References
[1]   White, D.G., Hudson, C., Maurer, J.J. and Ayers, S. (2000) Characterization of Chloramphinicol and Florfenical Resistance in Escherichia coli Associated with Bovine Diarrhea. Journal of Clinical Microbiology, 38, 4593-4598.

[2]   Javeed, I., Hafeez, R. and Anwar, M.S. (2011) Antibiotic Susceptibility Pattern of Bacterial Isolates from Patients Admitted to a Tertiary Care Hospital in Lahore. Biomedica, 27, 19-23.

[3]   John, E. and McGrowan, J.R. (2006) Resistance in Non-Fermenting Gram-Negative Bacteria: Multidrug Resistance to the Maximum. American Journal of Infection Control, 34, 29-37.
http://dx.doi.org/10.1016/j.ajic.2006.05.226

[4]   Fluit, A.C., Visser, M.R. and Schmitz, F.J. (2002) Molecular Detection of Antimicrobial Resistance. Journal of Clinical Microbiology, 14, 836-871.
http://dx.doi.org/10.1128/CMR.14.4.836-871.2001

[5]   Enoch, D.A., Birkett, C.I. and Ludlam, H.A. (2007) Non Fermentative Gram-Negative Bacteria. International Journal of Antimicrobial Agents, 3, 33-41.

[6]   Meenakumari, S., Verma, S., Absar, A. and Chaudhary, A. (2011) Antimicrobial Susceptibility Pattern of Clinical Isolates of Pseudomonas aeruginosa in an Indian Cardiac Hospital. International Journal of Engineering Science and Technology, 3, 7117-7124.

[7]   Lambert, P.A. (2002) Mechanisms of Antibiotic Resistance in Pseudomonas aeruginosa. Journal of the Royal Society of Medicine, 95, 22-26.

[8]   Poole, K. (2004) Efflux-Mediated Multiresistance in Gram-Negative Bacteria. Clinical Microbiology and Infection, 10, 12-26.

[9]   Merlo, C.A., Boyle, M.P., Diener, W.M., Marshall, B.C., Goss, C.H. and Lechtzin, N. (2007) Incidence and Risk Factors for Multiple Antibiotic-Resistant Pseudomonas aeruginosa in Cystic Fibrosis. Chest, 132, 562-568.
http://dx.doi.org/10.1378/chest.06-2888

[10]   Rojas, L., Vinuesa, T., Tubau, F., Truchero, C., Benzc, R. and Vinas, M. (2006) Integron Presence in a Multiresistant Morganella Morganii Isolate. International Journal of Antimicrobial Agents, 27, 505-512.
http://dx.doi.org/10.1016/j.ijantimicag.2006.01.006

[11]   Biendo, M., Thomas, D., Laurans, G., Daoudi, F.H., Canarelli, B., Rousseau, F. and Castelain, S. (2005) Molecular Diversity of Proteus mirabilis Isolates Producingextended-Spectrum Beta-Lactamases in a French University Hospital. Clinical Microbiology and Infection, 11, 395-401.
http://dx.doi.org/10.1111/j.1469-0691.2005.01147.x

[12]   Butt, T., Ahmad, R.N., Salman, M. and Kazmi, S.Y. (2005) Changing Trends in Drug Resistance among Typhoid Salmonellae in Rawalpindi, Pakistan. Eastern Mediterranean Health Journal, 11, 1038-1044.

[13]   Su, L.H., Wu, T.L., Chia, J.H., Chu, C., Kuo, A.J. and Chiu, C.H. (2005) Increasing Ceftriaxone Resistance in Salmonella Isolates from a University Hospital in Taiwan. Journal of Antimicrobial Chemotherapy, 55, 846-852.
http://dx.doi.org/10.1093/jac/dki116

[14]   Wain, J. and Kidgell, C. (2004) The Emergence of Multidrug Resistance to Antimicrobial Agents for the Treatment of Typhoid Fever. Transactions of the Royal Society of Tropical Medicine and Hygiene, 98, 423-430.
http://dx.doi.org/10.1016/j.trstmh.2003.10.015

[15]   Cheesbrough, M. (2000) District Laboratory Practice Manual in Tropical Countries Part 2. Cambridge University Press, Cambridge, 178-179.

[16]   Harley, J. (1990) Prescott L. Laboratory Exercises in Microbiology. Wm. C. Brown Publishers, 49-53.

[17]   Gloria, A., Cheryl, B., John, E., Richard, F., Joan, S.K., Tanja, P., Joy, W. and Scott, F.D. (2003) Manual for the Laboratory Identification and Antimicrobial Susceptibility Testing of Bacterial Pathogens of Public Health Importance in the Developing World. Centers for Disease Control and Prevention, Atlanta, Georgia, USA and World Health Organization: Department of Communicable Disease Surveillance and Response, 103-118.

[18]   Struve, C., Forestier, C. and Krogfelt, K.A. (2003) Application of a Novel Multi-Screening Signature-Tagged Mutagenesis Assay for Identification of Klebsiella pneumoniae Genes Essential in Colonization and Infection. Micro, 149, 167-176.
http://dx.doi.org/10.1099/mic.0.25833-0

[19]   Roshan, M., Ikram, A., Mirza, I.A., Malik, N., Abbasi, S.A. and Alizai, S.A. (2011) Susceptibility Pattern of Extended Spectrum B-Lactamase Producing Isolates in Various Clinical Specimens. Journal of the College of Physicians and Surgeons Pakistan, 21, 342-346.

[20]   Mahmod, A. (2000) Bacteriology of Surgical Site Infections and Antibiotic Susceptibility Pattern of the Isolates at a Tertiary Care Hospital in Karachi. Journal of Pakistan Medical Association, 50, 256-259.

[21]   Upgade, A., Prabhu, N., Gopi, V. and Soundararajan, N. (2012) Current Status of Antibiotic Resistant Non-Fermentative Gram Negative Bacilli among Nosocomial Infections. Advances in Science and Research, 3, 738-742.

[22]   Moniri, R., Mosayebi, Z., Movahedian, A.H. and Mousavi, G.A. (2005) Emergence of Multi-Drug-Resistant Pseudomonas aeruginosa Isolates in Neonatal Septicemia. Journal of Infectious Disease and Antimicrobial Agents, 22, 39-44.

[23]   Singla, N., Kaistha, N., Gulati, N. and Chander, J. (2010) Morganella morganii Could Be an Important Intensive Care Unit Pathogen. Indian Journal of Critical Care Medicine, 14, 154-155.
http://dx.doi.org/10.4103/0972-5229.74176

[24]   Falagas, M.E., Kavvadia, P.K., Mantadakis, E., Kofteridis, D.P., Bliziotis, I.A., Saloustros, E., Marak, S. and Samonis, G. (2006) Morganellamorganii Infections in a General Tertiary. Journal of Hospital Infection, 34, 315-321.

[25]   Lee, I.K. and Liu, J.W. (2006) Clinical Characteristics and Risk Factors for Mortality in Morganella morganii Bacteremia. Microbiology and Infectious Diseases Journals, 39, 328-334.

[26]   Nagshetty, K., Shivannavar, T.C. and Gaddad, S.M. (2010) Antimicrobial Susceptibility of Salmonella Typhi in India. The Journal of Infection in Developing Countries, 4, 70-73.

[27]   Muthu, G., Suresh, A., Sumathy, G. and Srivani, R. (2011) Studies on Antimicrobial Susceptibility Pattern of Salmonella Isolates from Chennai, India. Internships of Pharmaceuticals and Biological Science, 2, 435-442.

[28]   Romao, C.M.C.P.A, Faria, Y.N.D., Pereira, L.R. and Asensi, M.D. (2005) Susceptibility of Clinical Isolates of Multiresistant Pseudomonas aeruginosa to a Hospital Disinfectant and Molecular Typing. Mem Inst Oswaldo Cruz Rio de Janeiro, 100, 541-548.

[29]   Tripathi, P., Banerjee, G., Saxena, S., Gupta, M.K. and Ramteke, P.W. (2011) Antibiotic Resistance Pattern of Pseudomonas aeruginosa Isolated from Patients of Lower Respiratory Tract Infection. African Journal of Microbiology Research, 5, 2955-2959.

[30]   Manian, F.A., Meyer, L., Jenne, J., Owen, A. and Taff, T. (1996) Loss of Antimicrobial Susceptibility in Aerobic Gram-Negative Bacilli Repeatedly Isolated from Patients in Intensive Care Units. Infection Control and Hospital Epidemiology, 17, 222-226.
http://dx.doi.org/10.2307/30141024

[31]   Zehra, A., Naqvi, B.S., Bushra, R. and Ali, S.Q. (2010) Comparative Study on Resistance Pattern of Different Pathogens against Cefixime and Cefepime. Journal of Pharmaceutical Sciences, 3, 145-156.

[32]   Xiao, Y.H., Wang, J., Zhu, Y., Qi, H.M., Li, X.Y., Zhao, C.Y. and Xue, F. (2010) Mohnarin of 2008: Surveillance Results of National Bacterial Drug Resistance. Chinical Journal of Nosocom, 16, 6.

[33]   Sharma, S., Gupta, A. and Arora, A. (2012) Cefepime Tazobactam: A New β Lactam/β Lactamase Inhibitor Combination against ESBL Producing Gram Negative Bacilli. International Journal of Pharmaceutical and Biomedical Research, 3, 35-38.

[34]   Mushtaq, M.A. (2006) What after Ciprofloxacin and Ceftriaxone in Treatment of Salmonella typhi. Pakistan Journal of Medical Sciences, 22, 51-54.

[35]   Kumar, S., Rizv, M. and Berry, N. (2008) Rising Prevalence of Enteric Fever Due to Multidrug Resistant Salmonella: An Epidemiological Study. Journal of Medical Microbiology, 57, 1247-1250.
http://dx.doi.org/10.1099/jmm.0.2008/001719-0

[36]   Shankar, N., Baghdayan, A.S. and Gilmore, M.S. (2002) Modulation of Virulence within a Pathogenicity Island Vancomycin-Resistant Enterococcus feacalis. Nature, 417, 746-750.
http://dx.doi.org/10.1038/nature00802

[37]   Sava, G., Heikens, E. and Huebner, J. (2010) Pathogenesis and Immunity in Enterococcal Infections. Clinical Microbiology and Infection, 16, 533-540.
http://dx.doi.org/10.1111/j.1469-0691.2010.03213.x

 
 
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