Antibiotic susceptibility and phylogenetic analyses for the origins and serotypes of Listeria monocytogenes strains isolated from ice cream and cream cakes

Listeria monocytogenes is a zoonotic bacterium which also infects humans. The aim of this study was to isolate this organism from cream cakes and ice cream and obtain 16S rRNA and hlyA gene sequences from isolates in order to perform phylogenetic analyses. Serotypes and antibiotic susceptibility were also determined. The cream cake and ice cream samples were examined for L. monocytogenes according to ISO 11290-1 and using the mini Vidas LMO 2 kit procedure. Antibiotic susceptibilities were investigated using the disc diffusion method according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) and the Clinical and Laboratory Standards Institute (CLSI) standards. The Sanger DNA sequencing method for phylogenetic analysis was used for L. monocytogenes isolates. A total of 16 (n =128, 12.50%) L. monocytogenes strains, 9 (12.16%) from cream cake samples and 7 (12.96%) from ice cream samples, were isolated. This was the first investigation involving sequencing of L. monocytogenes isolated from cream cakes and ice creams in Turkey. All isolates were susceptible to sulfamethoxazole/trimethoprim, tetracycline, streptomycin, gentamicin, meropenem, and erythromycin. The numbers of isolates resistant to sulbactam/ampicillin, penicillin G, chloramphenicol, and ampicillin were 16, 2, 1, and 1, respectively. Moreover, 3 isolates showed intermediate resistance to amikacin and 2 to ciprofloxacin. The hlyA gene sequences of 11 of the L. monocytogenesisolates isolated from milk were closely related to the hlyA gene sequences of the GenBank reference strains. The comparison of the 16s rRNA gene sequences of the L. monocytogenes strains with the GenBank reference serotypes identified 1 isolate as serotype 1/2c, 1 as serotype 1/2a, and 1 as serotype 4. Nucleic acid sequencing is useful for identification of L. monocytogenes. The 16S rRNAand hlyA sequences can be used to determine the origin and relationship between L. monocytogenes isolates, as well as the serotype.

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1. Churchill KJ, Sargeant JM, Farber JM, O’connor AM. Prevalence of Listeria monocytogenes in select ready-to-eat foods—deli meat, soft cheese, and packaged salad: A systematic review and meta-analysis. Journal of Food Protection. 2019; 82 (2): 344- 357. doi:10.4315/0362-028X.JFP-18-158
2. Kocaman N, Sarımehmetoğlu B. Isolation of Listeria monocytogenes in lamb meat and determination of the antibiotic resistance. Ankara University Veterinary Faculty Journal. 2017; 64 (4): 273-279. doi:10.1501/vetfak_0000002810
3. Procop GW, Church DL, Hall GS, Janda WM, Koneman EW, et al. Koneman’s color atlas & textbook of diagnostic microbiology. 7th. ed. Philadelphia: Wolters Kluwer; 2017. pp. 2019-2035
4. Jagadeesan B, Bastic Schmid V, Kupski B, McMahon W, Klijn A. Detection of Listeria spp. and L. monocytogenes in pooled test portion samples of processed dairy products. International Journal of Food Microbiology 2019; 289 (August 2018): 30-39. doi:10.1016/j.ijfoodmicro.2018.08.017
5. Lee S, Cappato L, Guimarães J, Balthazar C, Rocha R, et al. Listeria monocytogenes in milk: occurrence and recent advances in methods for inactivation. Beverages 2019; 5(1): 14. doi:10.3390/beverages5010014
6. Lianou A, Moschonas G, Nychas GJE, Panagou EZ. Growth of Listeria monocytogenes in pasteurized vanilla cream pudding as affected by storage temperature and the presence of cinnamon extract. Food Reserach International 2018; 106 (November 2017): 1114-1122. doi:10.1016/j.foodres.2017.11.027
7. Buchanan RL, Gorris LGM, Hayman MM, Jackson TC, Whiting RC. A review of Listeria monocytogenes : An update on outbreaks, virulence, dose-response, ecology and risk assessments. Food Control 2017; 75: 1–13.
8. Singh A, Kour H, Dogra S, Dogra S. Contamination of dairy products by Listeria monocytogenes and its control. Imperial Journal of Interdisciplinary Research 2017; 1: 1293-1295.
9. Rahimi E, Ameri M, Momtaz H. Prevalence and antimicrobial resistance of Listeria species isolated from milk and dairy products in Iran. Food Control 2010; 21 (11): 1448-1452. doi:10.1016/j.foodcont.2010.03.014
10. Şanlıbaba P, Uymaz Tezel B, Çakmak GA. Detection of Listeria spp. in raw milk and dairy products retailed in Ankara. Journal of Food 2018; 43 (2): 273-282. doi:10.15237/gida.gd17107
11. Silbernagel KM, Carver CN, Jechorek RB, Johnson RL. Evaluation of VIDAS Listeria monocytogenes II (LMO2) immunoassay method for the detection of listeria monocytogenes in foods: collaborative study. Food Biological Contaminants 2004; 87 (5): 1123-1132.
12. Ligozzi M, Bernin C, Bonora MG, de Fatima M, Zuliani J, et al. Evaluation of the VITEK 2 system for identification and antimicrobial susceptibility testing of medically relevant grampositive cocci. Journal of Clinical Microbiology 2002; 40 (5): 1681-1686.
13. Woese CR. Bacterial evolution. Microbiology Reviews 1987; 51: 221–227.
14. Soni DK, Dubey SK. Phylogenetic analysis of Listeria monocytogenes based on sequencing of 16S rRNA and hlyA genes. Molecular Biology Reports 2014; 41 (12): 8219-8229. doi:10.1007/s11033-014-3724-2
15. Hellberg RS, Martin KG, Keys AL, Haney CJ, Shen Y, et al. 16S rRNA partial gene sequencing for the differentiation and molecular subtyping of Listeria species. Food Microbiology 2013; 36 (2): 231-240. doi:10.1016/j.fm.2013.06.001
16. Moreno LZ, Paixa˜o R de Gobbi DDS, Raimundo DC, Ferreira TSP, Moreno AK, et al. Phenotypic and genotypic characterization of atypical Listeria monocytogenes and Listeria innocua isolated from swine slaughterhouses and meat markets. Biomedical Research International 2014; 2014: 742032 doi:10.1155/2014/742032
17. Ricci A, Allende A, Bolton D, Chemaly M, Davies R, et al. Listeria monocytogenes contamination of ready-to-eat foods and the risk for human health in the EU. EFSA Journal. 2018; 16 (1): 5134. doi:10.2903/j.efsa.2018.5134
18. Hossain MK, Paul S, Hossain MM, Islam MR, Alam MGS. Bovine mastitis and its therapeutic strategy doing antibiotic sensitivity test. Austin Journal of Veterinary Science & Animal Husbandry 2017; 4 (1): 1030.
19. Anonymous. Turkish Food Codex Microbiological Criteria Rules. Official Gazette of the Republic of Turkey 2011; 28157 (3).
20. Abdollahzadeh E, Ojagh SM, Hosseini H, Irajian G, Ghaemi EA. Prevalence and molecular characterization of Listeria spp. and Listeria monocytogenes isolated from fish, shrimp, and cooked ready-to-eat (RTE) aquatic products in Iran. LwtFood Science and Technology 2016; 73: 205-211. doi:10.1016/j. lwt.2016.06.020
21. Arslan S, Özdemir F. Prevalence and antimicrobial resistance of Listeria spp. in homemade white cheese. Food Control 2008; 19 (4): 360-363. doi: 10.1016/j.foodcont. 2007.04.009
22. Kevenk TO, Terzi Gulel G. Prevalence, antimicrobial resistance and serotype distribution of Listeria monocytogenes isolated from raw milk and dairy products. Journal of Food Safety 2016; 36: 11-18. doi: 10.1111/jfs.12208
23. Sağun E, Sancak YC, İşleyici O, Ekici K. The presence and prevalence of Listeria species in milk and herby cheese in and around Van. Turkish Journal of Veterinary & Animal Sciences 2001; 25: 15-19.
24. Erol I, Şireli UT. Occurrence and contamination levels of Listeria spp. in milk and dairy products in Ankara. FEMS Symposium on the versatility of Listeria species 2002; 10: 11- 12.
25. International Standards Institute (ISO). 11290-1. Microbiology of the food chain — Horizontal method for the detection and enumeration of Listeria monocytogenes and of Listeria spp. — Part 1: Detection method. 2017.
26. Johnson R, Mills J. VIDAS® Listeria monocytogenes II (LMO2): GovVal validation comparison to the health canada mfhpb-30 reference method for detection of Listeria monocytogenes in ready-to-eat meats. Journal of AOA C International 2013; 96 (2): 246-250.
27. Bauer AW, Kirby WM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology 1966; 45: 493-496.
28. Clinical and Laboratory Standards Institute (CLSI). Performance standards for antimicrobial susceptibility testing; twenty second informational supplement (Vol. 32, No. 3). Wayne, PA, USA: Clinical and Laboratory Standards Institute, M100-S22. 2012.
29. Clinical and Laboratory Standards Institute (CLSI). Performance standards for antimicrobial susceptibility testing; twenty forth informational supplement (Vol. 34, No. 1). Wayne, PA, USA: Clinical and Laboratory Standards Institute, M100-S24. 2014.
30. European Committee on Antimicrobial Susceptibility Testing (EUCAST). (2019). Breakpoint tables for interpretation of MICs and zone diameters. Version 9.0.
31. Aksoy A, Sezer Ç, Vatansever L, Gülbaz G. Presence and antibiotic resistance of Listeria monocytogenes in raw milk and dairy products. Kafkas Üniversitesi Veteriner Fakültesi Dergisi 2018; 24 (3): 415-421. doi:10.9775/kvfd.2017.19081
32. Sanger F, Coulson AR. A rapid method for determining sequences in DNA by primed synthesis with DNA polymerase. Journal of Molecular Biology 1975; 94 (3): 441-448. doi:10.1016/0022-2836(75)90213-2
33. Saitou, N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution 1987; 4 (4): 406-425. doi:10.1093/oxfordjournals. molbev.a040454
34. Gönülalan S, Gönülalan Z. Detection of Listeria monocytogenes in ice cream samples retailed in Kayseri city of Turkey. Journal of Health Sciences 2010; 19 (3): 191-195.
35. Tahoun ABMB, Abou Elez RMM, Abdelfatah EN, Elsohaby I, El-Gedawy AA, et al. Listeria monocytogenes in raw milk, milking equipment and dairy workers: molecular characterization and antimicrobial resistance patterns. Journal of Global Antimicrobial Resistance. 2017; 10: 264-270. doi:10.1016/j.jgar.2017.07.008
36. Akrami-Mohajeri F, Derakhshan Z, Ferrante M, Hamidiyan N, Soleymani M, et al. The prevalence and antimicrobial resistance of Listeria spp in raw milk and traditional dairy products delivered in Yazd, central Iran (2016). Food and Chemical Toxicology 2018; 114: 141-144. doi:10.1016/j.fct.2018.02.006
37. Wijendra WAS, Kulathunga KACG, Ramesh R. First report of Listeria monocytogenes serotypes detected from milk and milk products in Sri Lanka. Advances in Animal and Veterinary Sciences 2014; 2: 11-16.
38. Mary MS, ND S. Pervasiveness of Listeria monocytogenes in milk and dairy products. Journal of Food: Microbiology, Safety & Hygiene 2017; 02 (03). doi.org/10.4172/2476-2059.1000125
39. Wang W, Arshad MI, Khurshid M, Rasool MH, Nisar MA, Aslam MA, Qamar MU. Antibiotic resistance : a rundown of a global crisis. Infection and Drug Resistance 2018; 11: 1645- 1658.
40. Bruce JL, Hubner RJ, Cole EM, McDowell CI, Webster JA. Sets of EcoRI fragments containing ribosomal RNA sequences are conserved among different strains of Listeria monocytogenes. Proceedings of the National Academy of Sciences 1995; 92: 5229-5234
41. Jeffers GT, Bruce JL, McDonough P, Scarlet J, Boor KJ, et al. Comparative genetic characterisation of Listeria monocytogenes isolates for human and animal listeriosis cases. Microbiology 2001; 147: 1095-1104.