Multiple drug resistance of Listeria monocytogenes isolated from aborted women by using serological and molecular techniques in Diwaniyah city/ Iraq
Background and Objectives: The study was sought to detect the effect of Listeria monocytogenes on pregnant Iraqi women at Al-Diwaniya hospitals and determination of virulence genes and antimicrobial susceptibility of isolates.
Materials and Methods: 360 specimens including blood, urine, vaginal and endocervical were collected from 90 patients with spontaneous abortions. Blood samples were displayed to immunological study and remaining specimens were subjected to bacteriological diagnosis. PCR was used to determine the virulence factors and antimicrobial resistance genes.
Results: Fifteen positive samples (16.6%) of patients and thirteen isolates (14.5%) from patients were recognized based on ELISA and PCR assay respectively. The general isolation of L. monocytogenes strains in cases of abortive women was 13/270 (4.8%). L. monocytogenes strains were highly virulent because of presence of virulence factors associated genes, namely actA, hlyA, plcA and prfA in all strains. Multiple drug resistance (MAR) index values of 15.4% of isolates were >0.2.
Conclusion: It is necessary for conducting susceptibility testing and to select the suitable antibiotics and avoid the effects of these bacteria in pregnant women.
2. Dhama K, Kumaragurubaran K, Ruchi T, Muhammad Z, Shabbir S, Barbuddhe S, et al. Listeriosis in animals, its public health significance (food-borne zoonosis) and advances in diagnosis and control: a comprehensive review. Vet Q 2015; 35: 211-235.
3. Allerberge F, Bagó Z, Huhulescu S, Pietzka A (2014). Listeriosis: The Dark Side of Refrigeration and Ensiling. In: Sing A. (eds) Zoonoses - Infections Affecting Humans and Animals; Springer, Dordrecht.
4. Martinez MR, Wiedmann M, Ferguson M, Datta AR. Assessment of Listeria monocytogenes virulence in the Galleria mellonella insect larvae model. PLoS One 2017; 12(9):e0184557.
5. Slama R, Bouyer J, Windham G, FensterL, Werwatz A, Swan S. Influence of paternal age on the risk of spontaneous abortion. Am J Epidemiol 2005; 161: 816-823.
6. Kureljušiæ J, Rokviæ N, Jezdimiroviæ N, Kureljušiæ B, Pisinov B, Karabasil N. Isolation and detection of Listeria monocytogenes in poultry meat by standard culture methods and PCR. IOP Conf Ser Earth Environ Sci 2017; 85: 012-069.
7. Lamont RF, Sobel J, Mazaki-Tovi S. 'Listeriosis in human pregnancy: a systematic review'. J Perinat Med 2011; 39: 227-236.
8. Goodwin PR. Food allergen detection methods: A coordinated approach. J AOAC Int 2004; 87: 1383-1390.
9. Rawool D, Malik S, Shakuntala I, Sahare A, Barbuddhe S. Detection of multiple virulence-associated genes in Listeria monocytogenes isolated from bovine mastitis cases. Int J Food Microbiol 2007; 113: 201-207.
10. Lotfollahi L, Nowrouzi J, Irajian G, Masjedian F, Kazemi B, Eslamian L, et al. Prevalence and antimicrobial resistence profiles of Listeria monocytogenes in spontaneous abortions in humans. Afr J Microbiol Res 2011; 5: 1990-1993.
11. Kaur S, Malik S, Vaidya V, Barbuddhe S. Listeria monocytogenes in spontaneous abortions in humans and its detection by multiplex PCR. J Appl Microbiol 2007; 103: 1889-1896.
12. Heidarzadeh S, Dallal SMM, Mohammad RP, Pirjani R, Foroushani RA, Noori M, et al. Prevalence, antimicrobial susceptibility, serotyping and virulence genes screening of Listeria monocytogenes strains at a tertiary care hospital in Tehran, Iran. Iran J Microbiol 2018; 10: 307-313.
13. National Committee for Clinical Laboratory Standards (2004). Procedures for the Handling and Processing of Blood Specimens; Approved Guideline-Third Edition. H18-A3. Wayne, Pennsylvania.
14. Ueda S, Kuwabara Y. Evaluation of an enzyme-linked fluorescent assay for the detection of Listeria monocytogenes from food. Biocontrol Sci 2010; 15: 91-95.
15. McLauchlin J, Rees CED (2009). Genus Listeria. In: De Vos P, Garrity G, Jones D, Krieg NR, Ludwig W, Rainey FA, Schleifer KH, Whitman WB. The low G + C Gram-positive bacteria. Bergey’s manual of systematic bacteriology. 2,3 Ed, Springer Dordrecht pp: 244-257.
16. Pospiech T, Neumann J (1995). In Genomic DNA isolation T. Kieser eds. John Innes Center. Norwich NR4 7UH.U.K.
17. Notermans SHW, Dufrenne J, Leimeister-Wachter M, Domann E, Chakraborty T. Phosphatidy-linositol-specific phospholipase C activity as a marker to distinguish between pathogenic and non-pathogenic Listeria species. Appl Environ Microbiol 1991; 57: 2666-2670.
18. Cooray KJ, Nishibori T, Xiong H, Matsuyama T, Fujita M, Mitsuyama M. Detection of multiple virulence-associated genes of Listeria monocytogenes by PCR in artificially contaminated milk samples. Appl Environ Microbiol 1994; 60: 3023-3026.
19. Paziak-Domańska B, Bogusławska E, Wieckowska-Szakiel M, Kotłowski R, Rózalska B, Chmiela M, et al. Evaluation of the API test, phosphatidylinositol-specific phospholipase C activity and PCR method in identification of Listeria monocytogenes in meat foods. FEMS Microbiol Lett 1999; 171: 209-214.
20. Suárez M, González-Zorn B, Vega Y, Chico-Calero I, Vázquez-Boland JA. A role for ActA in epithelial cell invasion by Listeria monocytogenes. Cell Microbiol 2001; 3: 853-864.
21. Kirby WMM, Bauer AW, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disc method. Am J Clin Pathol 1966; 45: 493-496.
22. European Committee on Antimicrobial Susceptibility Testin. Breakpoints tables for interpretation of MICs and zones diameters.2019. Version 9.0.
23. Krumpernam PH. Multiple antibiotic resistance indexing Escherichia coli to identify risk sources of fecal contamination of foods. Appl Environ Microbiol 1983; 46: 165-170.
24. Ikeh C, Obi SKC, Ezeasor D, Ezeonu IM, Moneke A. Incidence and pathogenicity profi le of Listeria spp. isolated from food and environmental samples in Nsukka, Nigeria. Afr J Biotechnol 2010; 9: 4776-4782.
25. Amagliani G, Omiccioli E, Campo A, Bruce IJ, Brandi G, Magnani M. Development of a magnetic capture hybridization-PCR assay for Listeria monocytogenes direct detection in milk samples. J Appl Microbiol 2006;100:375-383.
26. Yousif MG, AL-Shamari AK. Phylogenetinc characterization of Listeria monocytogenes isolated from different sources in Iraq. Asian J Pharm Clin Res 2018; 11:289-292.
27. Campbell S, Monga A (2000). Disorders of early pregnancy (ectopic miscarriage, gestataional trophoblastic disorders. In Gynaecology by Ten Teachers. 17th ed. pp: 102-104. Oxford University Press, Inc. New York.
28. Annas G (2007). Pregnancy loss. Obstetric: Normal and problem pregnancies. 5ed, Churchill living stone pp: 123.
29. Rezaei M, Pour NK, Vandyousefi J, Zamin FR, Irajian G. Determination of dominant serovars of Listeria monocytogenes strains isolated from spontaneous human abortion in Tehran/IRAN. Eurasia J Biosci 2018; 12: 377-383.
30. Shaker ME, Hassanien AA. PCR technique for detection of some virulence associated genes in Listeria monocytogenes isolated from table eggs and clinical human samples. Assiut Vet Med J 2015; 61: 219-225.
31. Park S, Szonyi B, Gautam R, Nightingale K, Anciso J, Ivanek R. Risk factors for microbial contamination in fruits and vegetables at the preharvest level: a systematic review. J Food Prot 2012; 75: 2055-2081.
32. Shakuntala I, Malik SVS, Barbuddhe SB, Rawool DB. Isolation of Listeria monocytogenes from buffaloes with reproductive disorders and its confirmation by polymerase chain reaction. Vet Microbiol 2006; 117: 229-234.
33. Chen J, Chen Q, Jiang J, Hu H, Ye J, Fang W. Serovar 4b complex predominates among Listeria monocytogenes isolates from imported aquatic products in China. Foodborne Pathog Dis 2010; 7: 31-41.
34. Caruso M, Fraccalvieri R, Pasquali F, Santagada G, Latorre LM, Difato LM, et al. Antimicrobial susceptibility and multilocus sequence typing of Listeria monocytogenes isolated over 11 years from food, humans, and the environment in Italy. Foodborne Pathog Dis 2020; 17: 284-294.