Abstract
Listeria monocytogenes, a life-threatening pathogen, poses severe risk during pregnancy, may cause abortion, fetal death or neonatal morbidity in terms of septicemia and meningitis. The present study aimed at characterizing L. monocytogenes isolated from pregnant women based on serotyping, antibiotic susceptibility, virulence genes, in vivo pathogenicity test and ERIC- and REP-PCR fingerprint analyses. The results revealed that out of 3700 human clinical samples, a total of 30 (0.81%) isolates [12 (0.80%) from placental bit (1500), 18 (0.81%) from vaginal swab (2200)] were positive for L. monocytogenes. All the isolates belonged to serogroup 4b, and were + ve for virulence genes tested i.e. inlA, inlC, inlJ, plcA, prfA, actA, hlyA, and iap. Based on the mice inoculation tests, 20 isolates showed 100% and 4 isolates 60% relative virulence while 6 isolates were non-pathogenic. Moreover, 2 and 10 isolates were resistant to ciprofloxacin and cefoxitin, respectively, while the rest susceptible to other antibiotics used in this study. ERIC- and REP-PCR collectively depicted that the isolates from placental bit and vaginal swab had distinct PCR fingerprints except a few isolates with identical patterns. This study demonstrates prevalence of pathogenic strains mostly resistant to cefoxitin and/or ciprofloxacin. The results indicate the importance of isolating and characterizing the pathogen from human clinical samples as the pre-requisite for accurate epidemiological investigations.
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Allerberger, F. and Wagner, M. 2010. Listeriosis: a resurgent foodborne infection. Clin. Microbiol. Infect. 16, 16–23.
Almeida, G., Morvan, A., Magalhaes, R., Santos, I., Hogg, T., Leclercq, A., Teixeira, P., and Research Team. 2010. Distribution and characterization of Listeria monocytogenes clinical isolates in Portugal, 1994-2007. Eur. J. Clin. Microbiol. Infect. Dis. 29, 1219–1227.
Anonymous. 1997. EN ISO 11290-1 Microbiology of food and animal feeding stuffs - Horizontal method for the detection and enumeration of Listeria monocytogenes - Part 1: Detection. International Organization for Standardisation, Geneva.
Aurora, R., Prakash, A., Prakash, S., Rawool, D.B., and Barbuddhe, S.B. 2008. Comparison of PI-PLC based assays and PCR along with in vivo pathogenicity tests for rapid detection of pathogenic Listeria monocytogenes. Food Control 19, 641–647.
Awofisayo, A., Amar, C., Ruggles, R., Elson, R., Adak, G.K., and Grant, K.A. 2015. Pregnancy-associated listeriosis in England and Wales. Epidemiol. Infect. 143, 249–256.
Bauer, A.W., Kirby, W.M., Sherris, J.C., and Turck, M. 1966. Antibiotic susceptibility testing by a standardized single disk method. Am. J. Clin. Pathol. 45, 493–496.
Camejo, A., Carvalho, F., Reis, O., Leitao, E., Sousa, S., and Cabanes, D. 2011. The arsenal of virulence factors deployed by Listeria monocytogenes to promote its cell infection cycle. Virulence 2, 379–394.
Chen, M., Wu, Q., Zhang, J., Yan, Z., and Wang, J. 2014. Prevalence and characterization of Listeria monocytogenes isolated from retail-level ready-to-eat foods in South China. Food Control 38, 1–7.
Chenal-Francisque, V., Diancourt, L., Cantinelli, T., Passet, V., Tran- Hykes, C., Bracq-Dieye, H., Leclercq, A., Pourcel, C., Lecuit, M., and Brisse, S. 2013. Optimized multilocus variable-number tendem repeat analysis assay and its complementarity with pulsedfield gel electrophoresis and multilocus sequence analysis for Listeria monocytogenes clone identification and surveillance. J. Clin. Microbiol. 51, 1868–1880.
Chou, C.H. and Wang, C. 2006. Genetic relatedness between Listeria monocytogenes isolates from seafood and humans using PFGE and REP-PCR. Int. J. Food Microbiol. 110, 135–148.
Clinical and Laboratory Standards Institute (CLSI). 2006 Performance standards for antimicrobial disk susceptibility tests; Approved standard-Ninth edition (M2-A9), Wayne, Pa, USA.
Conter, M., Paludi, D., Zanardi, E., Ghidini, S., Vergara, A., and Lanieri, A. 2009. Characterization of antimicrobial resistance of foodborne Listeria monocytogenes. Int. J. Food Microbiol. 128, 497–500.
Datta, A.R., Laksanalamai, P., and Solomotis, M. 2013. Recent development in molecular sub-typing of Listeria monocytogenes. Food Addit. Contam. A. 30, 1437–1445.
den Bakker, H.C., Warchocki, S., Wright, E.M., Allred, A.F., Ahlstrom, C., Manuel, C.S., Stasiewicz, M.J., Burrell, A., Roof, S., Strawn, L.K., et al. 2014. Listeria floridensis sp. nov., Listeria aquatica sp. nov., Listeria cornellensis sp. nov., Listeria riparia sp. nov. and Listeria grandensis sp. nov., from agricultural and natural environments. Int. J. Syst. Evol. Microbiol. 64, 1882–1889.
Dhanashree, B., Otta, S.K., Karunasagar, I., Goebel, W., and Karunasagar, I. 2003. Incidence of Listeria spp. in clinical and food samples in Mangalore, India. Food Microbiol. 20, 447–453.
Doumith, M., Buchrieser, C., Glaser, P., Jacquet, C., and Martin, P. 2004. Differentiation of the major Listeria monocytogenes serovars by multiplex PCR. J. Clin. Microbiol. 42, 3819–3822.
Fugett, E.B., Schoonmaker-Bopp, D., Dumas, N.B., Corby, J., and Wiedmann, M. 2007. Pulsed-field gel electrophoresis (PFGE) analysis of temporally matched Listeria monocytogenes isolates from human clinical cases, foods, ruminant farms, and urban and natural environments reveals source-associated as well as widely distributed PFGE types. J. Clin. Microbiol. 45, 865–873.
Goulet, V., Hebert, M., Hedberg, C., Laurent, E., Vaillant, V., De Valk, H., and Desenclos, J.C. 2012. Incidence of listeriosis and related mortalty among groups at risk of acquiring listeriosis. Clin. Infect. Dis. 54, 652–660.
Guillet, C., Join-Lambert, O., Le Monnier, A., Leclercq, A., Mechai, F., Mamzer-Bruneel, M.F., Bielecka, M.K., Scortti, M., Disson, O., Berche, P., et al. 2010. Human listeriosis caused by Listeria ivanovii. Emerg. Infect. Dis. 16, 136–138.
Huang, B., Fang, N., Dimovski, K., Wang, X., Hogg, G., and Bates, J. 2011 Observation of a new pattern in serogroup-related PCR typing of Listeria monocytogenes 4b isolates. J. Clin. Microbiol. 49, 426–429.
Jackson, K.A., Iwamoto, M., and Swerdlow, D. 2010. Pregnancy-associated listeriosis. Epidemiol. Infect. 138, 1503–1509.
Jamali, H. and Thong, K.L. 2014. Genotypic characterization and antimicrobial resistance of Listeria monocytogenes from ready-to-eat foods. Food Control 44, 1–6.
Kashish, Soni, D.K., Mishra, S.K., Prakash, R., and Dubey, S.K. 2015. Label-free impedimetric detection of Listeria mononcytogenes based on poly-5-carboxy indole modified ssDNA probe. J. Biotechnol. 200, 70–76.
Kaur, S., Malik, S.V.S., Bhilegaonkar, K.N., Vaidya, V.M., and Barbuddhe, S.B. 2010. Use of a phospholipase-C assay, in vivo pathogenicity assays and PCR in assessing the virulence of Listeria spp. Vet. J. 184, 366–370.
Kaur, S., Malik, S.V.S., Vaidya, V.M., and Barbuddhe, S.B. 2007. Listeria monocytogenes in spontaneous abortions in humans and its detection by multiplex PCR. J. Appl. Microbiol. 103, 1889–1896.
Lamont, F.R., Sobel, J., Mazaki-Tovi, S., Kusanovic, J.P., Vaisbuch, E., Kim, S.K., Uldbjerg, N., and Romero, R. 2011. Listeriosis in human pregnancy: a systematic review. J. Perinat. Med. 39, 227–236.
Liu, D., Lawrence, M.L., Austin, F.W., and Ainsworth, A.J. 2007a. A multiplex PCR for species- and virulence-specific determination of Listeria monocytogenes. J. Microbiol. Meth. 71, 133–140.
Liu, D., Lawrence, M.L., Ainsworth, A.J., and Austin, F.W. 2007b. Toward an improved laboratory definition of Listeria monocytogenes virulence. Int. J. Food Microbiol. 118, 101–115.
Lyon, S.A., Berrang, M.E., Fedorka-Cray, P.J., Fletcher, D.L., and Meinersmann, R.J. 2008. Antimicrobial resistance of Listeria monocytogenes isolated from a poultry further processing plant. Foodborne Pathog. Dis. 5, 253–259.
Mammina, C., Aleo, A., Romani, C., Pellissier, N., Nicoletti, P., Pecile, P., Nastasi, A., and Pontello, M.M. 2009. Characterization of Listeria monocytogenes isolates from human listeriosis cases in Italy. J. Clin. Microbiol. 47, 2925–2930.
Mammina, C., Parisi, A., Guaita, A., Aleo, A., Bonura, C., Nastasi, A., and Pontello, M. 2013. Enhanced surveillance of invasive listeriosis in the Lombardy region, Italy, in the year 2006–2010 reveals major clones and an increase in serotype 1/2a. BMC Infect. Dis. 13, 152.
Mateus, T., Silva, J., Maia, R.L., and Teixeira, P. 2013. Listeriosis during pregnancy: a public health concern. ISRN Obstet. Gynecol. 2013, 851712.
Menudier, A., Bosiraud, C., and Nicolas, J.A. 1991. Virulence of Listeria monocytogenes serovars and Listeria spp. in experimental infection of mice. J. Food Prot. 54, 917–921.
Mook, P., Grant, K.A., Little, C.L., Kafatos, G., and Gillespie, I.A. 2010. Emergence of pregnancy-related listeriosis amongst ethnic minorities in England and Wales. Euro. Surveill. 15, 17–23.
Morvan, A., Moubareck, C., Leclercq, A., Hervé-Bazin, M., Bremont, S., Lecuit, M., Courvalin, P., and Le Monnier, A. 2010. Antimicrobial resistance of Listeria monocytogenes strains isolated from humans in France. Antimicrob. Agents Chemother. 54, 2728–2731.
Nightingale, K.K., Windham, K., and Wiedmann, M. 2005. Evolution and molecular phylogeny of Listeria monocytogenes isolated from human and animal listeriosis cases and foods. J. Bacteriol. 187, 5537–5551.
Notermans, S.H., Dufrenne, J., Leimeister-Wachter, M., Domann, E., and Chakraborty, T. 1991. Phosphatidylinositol-specific phospholipase C activity as a marker to distinguish between pathogenic and non-pathogenic Listeria species. Appl. Environ. Microbiol. 57, 2666–2670.
Nwachukwu, N.C., Orji, F.A., Iheukwumere, I., and Ekeleme, U.G. 2010. Antibiotic resistant environmental isolates of Listeria monocytogenes from anthropogenic lakes in Lokpa-Ukwu, Abia State of Nigeria. Aust. J. Basic Appl. Sci. 4, 1571–1576.
Orsi, R.H., Ripoll, D.R., Yeung, M., Nightingale, K.K., and Wiedmann, M. 2007. Recombination and positive selection contribute to evolution of Listeria monocytogenes inlA. Microbiology 153, 2666–2678.
Poulsen, K.P. and Czuprynski, C.J. 2013. Pathogenesis of listeriosis during pregnancy. Anim. Health Res. Rev. 14, 30–39.
Rivera, I.G., Chowdhury, M.A.R., Huq, A., Jacobs, D., Martins, M.T., and Colwell, R.R. 1995. Enterobacterial repetitive intergenic consensus sequences and the PCR to generate fingerprints of genomic DNA from Vibrio cholerae O1, O139 and non-O1 strains. Appl. Environ. Microbiol. 61, 2898–2904.
Roche, S.M., Gracieux, P., Milohanic, E., Albert, I., Virlogeux-Payant, I., Temoin, S., Grepinet, O., Jacquet, C., Cossart, P., and Velge, P. 2005. Investigation of specific substitutions in virulence genes characterizing phenotypic groups of low-virulence field strains of Listeria monocytogenes. Appl. Environ. Microbiol. 71, 6039–6048.
Rodas-Suarez, O.R., Flores-Pedroche, J.F., Betancourt-Rule, J.M., Quinones-Ramirez, E.I., and Vazquez-Salinas, C. 2006. Occurrence and antibiotic sensitivity of Listeria monocytogenes strains isolated from oysters, fish and estuarine water. Appl. Environ. Microbiol. 72, 7410–7412.
Scallan, E., Hoekstra, R.M., Angulo, F.J., Tauxe, R.V., Widdowson, M.A., Roy, S.L., and Griffin, P.M. 2011. Foodborne illness acquired in the United States-major pathogens. Emerg. Infect. Dis. 17, 7–15.
Sharma, D., Sharma, P.K., Saharan, B.S., and Malik, A. 2012. Isolation, identification and antibiotic susceptibility profiling of antimicrobial resistant Listeria monocytogenes from dairy milk. Int. J. Microbial Resource Technol. 1, 1–4.
Shen, J., Rumpb, L., Zhang, Y., Chen, Y., Wang, X., and Meng, J. 2013. Molecular subtyping and virulence gene analysis of Listeria monocytogenes isolates from food. Food Microbiol. 35, 58–64.
Shi, W., Qingping, W., Jumei, Z., Moutong, C., and Zean, Y. 2015. Prevalence, antibiotic resistance and genetic diversity of Listeria monocytogenes isolated from retail ready-to-eat foods in China. Food Control 47, 340–347.
Soni, D.K. and Dubey, S.K. 2014. Phylogenetic analysis of the Listeria monocytogenes based on sequencing of 16S rRNA and hlyA genes. Mol. Biol. Rep. 41, 8219–8229.
Soni, D.K., Singh, K.M., Ghosh, A., Chikara, S.K., Joshi, C.G., and Dubey, S.K. 2015. Whole-genome sequence of Listeria monocytogenes strains from clinical and environmental samples from Varanasi, India. Genome Announc. 3, e01496-14.
Soni, D.K., Singh, R.K., Singh, D.V., and Dubey, S.K. 2013. Characterization of Listeria monocytogenes isolated from Ganges water, human clinical and milk samples at Varanasi, India. Infect. Genet. Evol. 14, 83–91.
Soni, D.K., Singh, M., Singh, D.V., and Dubey, S.K. 2014. Virulence and genotypic characterization of Listeria monocytogenes isolated from vegetable and soil samples. BMC Microbiol. 14, 241.
Takeuchi, K., Mytle, N., Lambert, S., Coleman, M., Doyle, M.P., and Smith, M.A. 2006. Comparison of Listeria monocytogenes virulence in a mouse model. J. Food Protect. 69, 842–846.
Van Stelten, A., Simpson, J.M., Ward, T.J., and Nightingale, K.K. 2010. Revelation by single-nucleotide polymorphism genotyping that mutations leading to a premature stop codon in inlA are common among Listeria monocytogenes isolates from ready-toeat foods but not human listeriosis cases. Appl. Environ. Microbiol. 76, 2783–2790.
Versalovic, J.T., Koeuth, T., and Lupski, J.R. 1991. Distribution of repetitive DNA sequences in Eubacteria and application to fingerprinting of bacterial genomes. Nucleic Acids Res. 19, 6823–6831.
Weller, D., Andrus, A., Wiedmann, M., and den Bakker, H.C. 2015. Listeria booriae sp. nov. and Listeria newyorkensis sp. nov., from food processing environments in the USA. Int. J. Syst. Evol. Microbiol. 65, 286–292.
World Health Organisation (WHO). 2004. Risk assessment of Listeria monocytogenes in ready-to-eat foods. Microbiological Risk Assessment Series 4, 1–78.
Yan, H., Neogi, S.B., Mo, Z., Guan, W., Shen, Z., Zhang, S., Li, L., Yamasaki, S., Shi, L., and Zhong, N. 2010. Prevalence and characterization of antimicrobial resistance of foodborne Listeria monocytogenes isolates in Hebei province of Northern China, 2005–2007. Int. J. Food Microbiol. 144, 310–316.
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Soni, D.K., Singh, D.V. & Dubey, S.K. Pregnancy - associated human listeriosis: Virulence and genotypic analysis of Listeria monocytogenes from clinical samples. J Microbiol. 53, 653–660 (2015). https://doi.org/10.1007/s12275-015-5243-9
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DOI: https://doi.org/10.1007/s12275-015-5243-9