Isolation and Identification of Burkholderia mallei and its Susceptibility to Anethum graveolens Extracts

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Khadija Khalil Mustafa


B. mallei Anethum graveolens Antimicrobial resistant.


In the present study, 13 (4.06%) isolates of Burkholderia mallei were obtained from  hands of 320 workers in veterinary laboratories in Erbil Province- Iraq. All isolates identified depending on conventional assays, VITEK 2 compact system, and further confirmed by PCR using 23 rDNA gen. The results of PCR product showed that the expected size (526bp) was obtained for all local isolates and for B. mallei ATCC 15310. Also the effect of 10 antimicrobials against isolated bacteria were studied and the results showed that isolated B. mallei were absolutely resistant to norfloxacin, and 100% sensitive to imipenem and ceftazidime. The antibacterial activity of aqueous and alcoholic extracts of Anethum  graveolens against B. mallei was studied. The results of MIC by using broth dilution method revealed that the alcohol and aqueous extracts exhibited inhibition of B. mallei growth and the MIC was 800, 1000 µg/ml, respectively. On molecular study, the SubMIC (750, 950 µg/ml) for both extracts against plasmid DNA profile of most resistant isolate of B. mallei was studied. The results showed three plasmid DNA band exist in local isolate of B. mallei and stander strain of B. mallei ATCC 15310 before treating with A. graveolens extracts.  While it was decreased from three to two in the local isolate and the standard strain ATCC 15310 when treated with A.  graveolens extracts. The inhibitory effect of the above mentioned extracts against total protein was studied by SDS-PAGE binding pattern. The results showed that there were differences in the protein banding pattern among the tested isolates and induction of some new protein bands when treated with plant extracts.

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ARORA, D. S. and KAUR, G. J. 2007. Antibacterial Activity of Some Indian Medicinal Plants. J Nat. Med. 61. P.313- 317.
BAESHIN, N. A.; TWATY, N. and Al-HEBSHI A. . 2005. Evaluation the genotoxicity of Rhazya stricta leaves extract by the Saccharomyces cerveisiae auxotrophic mutats test. Egyp. J. Nat. Toxi. 2. P.87-100.
BAILER, J. ; AICHINGER, T.; HACKI ,l. G.; HUEBER K. D. and DACHLER, M. 2001. Essential oil content and composition in commercially available dill cultivars in comparison to caraway. Indus Crops Prods. 14. P. 229-39.
BAUERNFEIND, A.; ROLLER, C.; MEYER, D., JUNGWIRTH, R. and SCHNEIDER, I. .1998. Molecular Procedure for Rapid Detection of Burkholderia mallei and Burkholderia pseudomallei. J. Clin Microbiol. 36(9).P. 2737–2741.
BURNS, J. L. Antibiotic resistance of Burkholderiaspp. In: Coenye T and Vandamme P, 2006. Burkholderia Molecular Microbiology and Genomic: 81–91. Horizon Bioscience. Wymondham. Norflok, UK .
CHAHAL, K.; MONIKA; KUMAR, A.; BHARDWAJ, U. and KAUR, R. 2017. Chemistry and biological activities of Anethum graveolensL. (dill) essential oil. J Pharmacognosy and Phytoch. 6(2). P. 295-306.
DAHIYA, P. and PURKAYASTHA , S. 2012. Phytochemical analysis and antibacterial efficacy of dill seed oil against multi-drug resistant clinical isolates. Asian Journal of Pharmaceutical and Clinical Research. 5(2).P. 62-64.
DANCE, D.; WUTHIEKANUN, V.; CHAOWAGUL, W. AND et al. 1989. The antimicrobial susceptibility of Pseudomonas pseudomallei. Emergence of resistance in vitro and during treatment.Journal of Antimicrobial Chemotherapy. 24, PP. 295–309.
CLINICAL and LABORATORY STANDARD INSTITUTE. 2011. Performance Standards for Antimicrobial Susceptibility Testing; Twenty-First Informational Supplement Clinical and Laboratory Standards Institute antimicrobial susceptibility testing standards, CLSI 31- M100-S16.
DELAQUIS, P. J.; STANICH, k.; GIRARD, B. and MAZZA, G. 2002. Antimicrobial activity of individual and mixed fractions of dill, cilantro, coriander and eucalyptus essential oils. J Food Microb. 74. P.101- 109.
DUKE, J. A. 2001. Handbooke of Medicinal Herbs. London: CRC Press, P. 42.
DVORAK, G. D. ; SPICKLER, A. R. and GLANDERS. J. 2008. American Vet. Med. Assoc. 233. P. 570–577
El-TARRAS, A. A.; HASSAN, M. M. and El-AWADY, M. 2013. Evaluation of the genetic effects of the in vitro antimicrobial activities of Rhazya stricta leaf extract using molecular techniques and scanning electron microscope, African J Biotechnology. 12(21).P. 3171-3180, DOI: 10.5897/AJB12.2467. ISSN 1684-5315.
GARVEY, M.; RAHMAN, M.; GIBBONS, S. and PIDDOCK, L. 2011. “Medicinal plant extracts with efflux inhibitory activity against Gram-negative bacteria, International J Antimicrobial Agents. 37( 2). P. 145–151.
HEINE, H. S.; ENGLAND, M. J.; WAAG, D. M. AND et al. 2001. In vitro antibiotic susceptibilities of Burkholderia mallei(causative agent of glanders) determined by broth microdilution and E-test. Antimicrobial Agents and Chemotherapy. 45. P. 2119–21.
HORN, J. K. 2003. Bacterial agents used for bioterrorism. Surg Infect (Larchmt), 4(3), PP. 281-287.
MAHDI, N. M. 2016. Antibacterial Effect of Dill Seed Oil Anethum graveolens Anethum graveolens Kufa J Nursing Sciences, 6( 2) .
MANDELL, G. B. and DOLIN, J. R., 2006. Pseudomonas species (including melioidosis and glanders). In Principles and practices of infectious disease 4th edition. Edited by: Mandell GBJ,Dolin R. New York: Churchill Livingstone.
MUHAMMAD, G.; KHAN, M. Z. and ATHAR, M. 1998. Clinico-microbiological and therapeutic aspects of glanders in equines. J. Equine Sci. 9 P. 93– 96.
MUSTAFA, KH. KH. 2007. Effect of medicinal plants on the growth of P. shigelloides isolated from different water sources, Duhok Scientific Journal. 3.
MUSTAFA, KH. KH.; MAULUD, S. Q. and HAMAD, P. A. 2017. Detection of Sphingomonas paucimobilisand antibacterial activity of Prosopis farctaextracts on it. Karbala International Journal of Modern Science: 1-7.Under publishing.
MUSTAFA, KH . KH and AL-RAWI . 2012. Antibacterial Activity of the Some Medicinal Plant Extract on the Growth of Legionella pneumophila Isolated From Different Water Sources. Journal of Koya University , No.23, 2012.
NAUREEN, A.; SAQIB, M.; MUHAMMAD, G.; HUSSAIN, M.H. and ASI, M.N. 2007. Comparative evaluation of Rose Bengal plate agglutination test, mullein and some conventional serological tests for diagnosis of equine glanders. J. Vet. Diag. Invest. 19. P. 362–367.
OIE. 2008. Manual of Diagnostic Tests and Vaccines for Terrestrial Animals, 8th edition,. Office International Des Epizooties. Paris. P. 919–928.
ORHAN, P. E.; SENOL, F. S., OZTURK, N.; CELIK, S. A. ; PULUR, A. and KAN, Y. 2003. Phytochemical contents and enzyme inhibitory and antioxidant properties of Anethum graveolens L. (dill) samples cultivated under organic and conventional agricultural conditions. J Food and Toxicology. 59. P. 96-103.
PATHAK, V.; DWIVEDI, R. and SHUKLA, P. 2014. Pharmacognostical study of Anethum sowa (dill) seed. International J Recent Biotechnology. 2(3). P. 6-14.
RAKHOLIYA, K. and CHANDA, S., 2012. “In vitro interaction of certain antimicrobial agents in combination with plant extracts against some pathogenic bacterial strains,” Asian Pacific Journal of Tropical Biomedicine. 2( 2) .P. S876–S880,
ROTZ, L. D.; KHAN, A. S.; LILLIBRDGE, S. R.; ASTROFF, S. M. and HUGHES, J. M. 2002. Public health assessment ofpotential biological terrorism agents. Emerg. Inf. Dis. 2. P. 225–230.
SAMBROOK, J. and RUSSELL, D. W. 2001. Molecular cloning. A laboratory manual, 3rd ed. New York: Cold Spring Harbor Laboratory Press.
SAQIB, M.;, MUHAMMAD, G.; NAUREEN, A.; SHAKOOR, A. and REHMAN, S. U. 2010. Disc diffusion based in vitro antibiotic susceptibility of recent isolates of Burkholderia mallei. Int. J. Agric. Biol. 12. P. 777–780 .
SAEI-DEHKORDI, S.; KHALIGHI-SIGIGAROODI, F.; PIRALKHEIRABADI, K.; ALIMARDANINAGHANI, F. and FALLAH, A. 2014. Chemical composition, antioxidative capacity and antimicrobial activity of Zeravschaina membranacea (boiss.) Pimenov essential oil. J Food Safety. 34: 87 - 94
THIBAULT, F. M.; HERNANDEZ, E.; VIDAL, D. R.; GIRARDET, M. and CAVALLO, J. D. 2004. Antibiotic susceptibility of 65 isolates of Burkholderia pseudomallei and Burkholderia mallei to 35 antimicrobial agents. J Antimicrobial Chemotherapy. 54. P. 1134–1138.
TRIBUDDARAT, C.; MOORE, R. A.; BAKER, P. and WOODS, D. E. 2003. Burkholderia pseudomallei class a beta-lactamase mutations that confer selective resistance against ceftazidime or clavulanic acid inhibition. Antimicrob. Agents Chemother. 47.P. 2082–2087.
WHITLOCK, G. C.; ESTES, D. M. and TORRES, A. G. 2007. Glanders: off to the races with Burkholderia mallei. FEMS Microbiol Lett. 277(2). P.115-122.