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Detection of Plasmid in Listeria monocytogenes Isolated from Fresh Raw Milk in Nigeria

Received: 2 October 2014     Accepted: 15 October 2014     Published: 30 October 2014
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Abstract

Aims: To determine the presence of plasmids in Listeria monocytogenes strains isolated from fresh raw milk from healthy cattle in Nigeria and also determine its role in antibiotics resistance since this organism is a food borne pathogen of public health importance. Methods and results: Isolation of Listeria monocytogenes was done using selective enrichment and isolation protocol, as recommended by United States Department of Agriculture (USDA) and Association of Analytical Chemists/ International Dairy Federation (AOAC/ IDF). Identification was by phenotypic characterization and confirmed using β-haemolytic activity, phosphatidyl inositol phospholipase-C (PIPLC) assay and polymerase chain reaction (PCR) . Strains identified as Listeria monocytogenes from the six zones were determined and results expressed as rate of prevalence in graph. Twenty-three strains of Listeria monocytogenes isolated from 305 samples of fresh raw milk were screened for the presence of plasmid DNA using the method of Ehrenfeld and Clewel and agarose gel electrophoresis. One isolate was found to harbour one plasmid with molecular weight of 14.62kb. The plasmid was cured using different concentrations of acridine orange. Antibiotic sensitivity test was carried out on both the wild strain (with plasmid) and cured strain using disc diffusion method of Bauer-Kirby and it was found that the wild strain was resistant to seven of the twelve antibiotics used while the cured strain was resistant to only one. Conclusions: There is a possibility of the presence of strains of Listeria monocytogenes that can harbour plasmids. This could confer antibiotic resistance to the strains and thus make antibiotic therapy unsuccessful. The increase in the number of antibiotics which the cured strain of Listeria monocytogenes was susceptible to in this study showed that plasmid may play a significant role in antibiotics resistance. The Fulani herdsmen who take fresh raw milk without boiling or pasteurization may serve as carriers of resistant strains of Listeria monocytogenes and help in its spread in the environment through faecal pollution.

Published in Animal and Veterinary Sciences (Volume 2, Issue 6)
DOI 10.11648/j.avs.20140206.12
Page(s) 169-174
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2014. Published by Science Publishing Group

Keywords

Fresh Raw Milk, Listeria monocytogenes, Plasmid, Antibiotic Resistance

References
[1] Low JC and Donachie W,(1997) A review of Listeria monocytogenes and listeriosis. Vet. J 153: 9-29.
[2] Zhang Y, Yeh E, Hall G, Crioe J,Bhaowat A.A, Meno J. (2007) Characterization of Listeria monocytogenes isolated from retail foods. Intl J. Food Microbiol. . 113 : 47-53
[3] Halter EL, Neuhaus K and Scherer S.(2012). Listeia weihenstephanensis sp. nov. isolated from the water plant Lemna trisulca of a German fresg water pond. Int. J. Syst. Evol. Microbiol. 63: 641-647.
[4] Liu D. (2006) Identification, subtyping and virulence determination of Listeria monocytogenes an important foodborne pathogen. J. Med.Microbiol. 55: 645-659
[5] Kuhn M, Scortti M and Vazquez-Boland JA (2008). Pathogenesis. In Liu, D (ed) Handbook of Listeria monocytogenes, CRC Press, New York. Pp 99-102.
[6] Liu, D (2008). Epidemioogy. In Liu, D (ed) Handbook of Listeria monocytogenes, CRC Press, New York, pp27-30.
[7] Mead PS, Slustker L, Dietz V and Bresee J. (1998). Food epidemic relaed illness ad the death in the United States. Emerg. Infect. Dis. 5 (5): 607-625.
[8] Lindback T, Secic I and Rorvik LM (2011). A contingency locus in prfA in a Listeria monocytogenes subgroup allows reactivation of the prfA virulence regulator during infection in mice. Appl. Environ. Microbiol. 77: 3478-3483.
[9] Chukwu OOC, Ogbonna CIC, Olabode AO, Chukwu AI, Onwuliri FC and Nwakiti OO (2006). Listeria monocytogenes in Nigerian processed mea and ready-to-eat-dairy products. Nigerian Journal of Microbiology 20 (2): 900-904.
[10] Enurah L.U., Aboaba O.O,Nwachukwu S.C.U, Nwosuh C.I. (2013) Antibiotic resistant profiles of food (fresh ra milk) and environmental (abattoir effluents) isolates of Listeria monocytogenes from the six zones of Nigeria. African Journal of Microbiology Research. 7 (34) 4373 4378.
[11] Salihu MD, Junaidu AU, Manga SB, Gulumbe ML, Magaji AA, Ahmed A, Adamu AY, Shittu A and Balarabe I (2008). Occurrence of Listeria monocytogenes in smoked fish in Sokoto, Nigeria. Afr. J. Biotechnol. 7: 3082-3084.
[12] Ikeh MAC, Obi SKC, Ezeasor DN, Ezeonu IM and Moneke AN (2010). Incidence and pathogenicity profile of Listeria sp. isolated from food and environmental samples in Nsukka, Nigeria. Afri. J. Biotechnol. 9 (30): 4776-4782.
[13] Clewell D.B. (1972) Nature of ColE1 plasmid replication in Escherichia coli in the presence of chloramphenicol. J. Bacteriol. 110. 667-676.
[14] Funnell B.E, Phillips G.J., editors,(2004) Plasmid Biology. ASM Press, Washington. DC, USA.
[15] USDA United States Department of Agriculture (1999) Listeria and food safety. Tips USD Consumer Publication List
[16] AOAC : Association of Analytical Chemists-International Methods of Analysis. 1995, 16th Edition
[17] IDF International Dairy Federation. Milk and milk products. Detection of Listeria monocytogenes. 43A, 1995. Hh//www.org/standardsEnglishhtw
[18] Wiedmann M, Bruce JL, Keating C, Johnson AE, McDonough PL and Batt CA.(1997) Ribotypes and virulence gene polymorphisms suggest three distinct Listeria monocytogenes. J. Bacteriol. 180: 1350- 1356.
[19] Ehrenfeld EE and Clewell DB.(1987) Methods in microbiology. J. Bacterial. 169: 3473-3481
[20] Meyers JA, Sanchez D, Elwell LP and Falkow S.(1976) Simple agarose gel electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid. J. Bacteriol. 127: 1529 – 1537.
[21] Valery Terletskry, Valentina Tyshchenko, Hargi Martinez-Ballesteros, Javier Garaizai and Joseba Bikandi. (2010)Validation of double digest selective label database for sequenced prokaryotic genomes. Bioinformatcs. 26 (3) 417-418
[22] CLSI. (2005) Performance standard for antimicrobial susceptibility testing, fifteenth informational supplement, M100-S15, vol.25. No. 1. Clinical and Laboratry Standard Institute , Wayne Pa
[23] Prentice GA.(1997) The significance of pathogenic microorganisms in raw milk, Listeria monocytogenes. Mijekartvo. 47: 147-162.
[24] Onyemelukwe GC, Lawande RV, Egler LJ and Mohammed I (1983). Listeria monocytogenes in Northern Nigeria. J. Infect. Dis. 6: 141-145
[25] Aurora R, Parakash A, Parakash S, Rawool DB and Barbuddhe SB. (2008) Comparison of PILC based assay and PCR along with invivo pathogenicity tests for rapid detection of pathogenic Listeria monocytogenes. Food Cont. 19: 641 – 647.
[26] Bhilegonkar KN, Killshrestha SB, Kapoor KN, Ashok- Kumar, Rgarwal RK, Singh BR and Kumar A.(1997) Isolatio of Listeria monocytogenes from milk. Int. J. Food Sci. and Tech. Mysore 34: 248 – 250.
[27] Sanaa M. Listeriosis: contamination of milk and milk products. Summa. 1996, 13: 65-75.
[28] Pesavento G, Doci B, Nieri O, Comodo N and Lo Nostro A.(2010) Prevalevce and antibiotic susceptibility of Listeria spp. isolated from raw meat and retail foods. Food Control. 21: 708-713.
[29] Pourshaban M, Ferrini A, Cannoni V, Oliva B and Aureli P. (2002)Transferable tetracycline resistance in Listeria monocytogenes from food in Italy. J. Med. Microbiol. 51 (suppl 7). 564-566.
[30] David OM and Odeyemi AT. (2007) Antibiotic resistant pattern of environmental isolates of Listeria monocytogenes from Ado- Ekiti, Nigeria. African J. Biotech. 6 (18): 2135- 2139
[31] Chao G, Zhou X, Jiao X, Qian X and Xu L. (2007) Prevalence and antimicrobial resistance of foodborne pathogens isolated from food products in China. Fooborne Pth. Dis. 4 (3): 277-284.
[32] Charpentier E, Gerbaud G, Jacquet C, Rocourt J and Courvalin P. (1995) Incidence of antibiotic resistance in Listeria spp. J. Infect Dis. 172: 277-281.
[33] Dancer SJ, Shears P and Platt D. (1997) Isolation and characterization of coliforms from glacial ice and water in Canadas High Artic. J. Appl. Microbiol. 82: 597-609.
[34] Finch RG. Antibiotic resistance. J. Antimicrob. Chem. (1998) 42:125-128
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    Enurah Leonard Uwanibe, Aboaba Olusimbo Olugbo, Nwachukwu Simon Cyril Ugwumba, Smith Stella Ifeanyi, Fowora Adenike, et al. (2014). Detection of Plasmid in Listeria monocytogenes Isolated from Fresh Raw Milk in Nigeria. Animal and Veterinary Sciences, 2(6), 169-174. https://doi.org/10.11648/j.avs.20140206.12

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    ACS Style

    Enurah Leonard Uwanibe; Aboaba Olusimbo Olugbo; Nwachukwu Simon Cyril Ugwumba; Smith Stella Ifeanyi; Fowora Adenike, et al. Detection of Plasmid in Listeria monocytogenes Isolated from Fresh Raw Milk in Nigeria. Anim. Vet. Sci. 2014, 2(6), 169-174. doi: 10.11648/j.avs.20140206.12

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    AMA Style

    Enurah Leonard Uwanibe, Aboaba Olusimbo Olugbo, Nwachukwu Simon Cyril Ugwumba, Smith Stella Ifeanyi, Fowora Adenike, et al. Detection of Plasmid in Listeria monocytogenes Isolated from Fresh Raw Milk in Nigeria. Anim Vet Sci. 2014;2(6):169-174. doi: 10.11648/j.avs.20140206.12

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  • @article{10.11648/j.avs.20140206.12,
      author = {Enurah Leonard Uwanibe and Aboaba Olusimbo Olugbo and Nwachukwu Simon Cyril Ugwumba and Smith Stella Ifeanyi and Fowora Adenike and Nwosuh Chika Ignatius},
      title = {Detection of Plasmid in Listeria monocytogenes Isolated from Fresh Raw Milk in Nigeria},
      journal = {Animal and Veterinary Sciences},
      volume = {2},
      number = {6},
      pages = {169-174},
      doi = {10.11648/j.avs.20140206.12},
      url = {https://doi.org/10.11648/j.avs.20140206.12},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.avs.20140206.12},
      abstract = {Aims: To determine the presence of plasmids in Listeria monocytogenes strains isolated from fresh raw milk from healthy cattle in Nigeria and also determine its role in antibiotics resistance since this organism is a food borne pathogen of public health importance. Methods and results: Isolation of Listeria monocytogenes was done using selective enrichment and isolation protocol, as recommended by United States Department of Agriculture (USDA) and Association of Analytical Chemists/ International Dairy Federation (AOAC/ IDF). Identification was by phenotypic characterization and confirmed using β-haemolytic activity, phosphatidyl inositol phospholipase-C (PIPLC) assay and polymerase chain reaction (PCR) . Strains identified as Listeria monocytogenes from the six zones were determined and results expressed as rate of prevalence in graph. Twenty-three strains of Listeria monocytogenes isolated from 305 samples of fresh raw milk were screened for the presence of plasmid DNA using the method of Ehrenfeld and Clewel and agarose gel electrophoresis. One isolate was found to harbour one plasmid with molecular weight of 14.62kb. The plasmid was cured using different concentrations of acridine orange. Antibiotic sensitivity test was carried out on both the wild strain (with plasmid) and cured strain using disc diffusion method of Bauer-Kirby and it was found that the wild strain was resistant to seven of the twelve antibiotics used while the cured strain was resistant to only one. Conclusions: There is a possibility of the presence of strains of Listeria monocytogenes that can harbour plasmids. This could confer antibiotic resistance to the strains and thus make antibiotic therapy unsuccessful. The increase in the number of antibiotics which the cured strain of Listeria monocytogenes was susceptible to in this study showed that plasmid may play a significant role in antibiotics resistance. The Fulani herdsmen who take fresh raw milk without boiling or pasteurization may serve as carriers of resistant strains of Listeria monocytogenes and help in its spread in the environment through faecal pollution.},
     year = {2014}
    }
    

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  • TY  - JOUR
    T1  - Detection of Plasmid in Listeria monocytogenes Isolated from Fresh Raw Milk in Nigeria
    AU  - Enurah Leonard Uwanibe
    AU  - Aboaba Olusimbo Olugbo
    AU  - Nwachukwu Simon Cyril Ugwumba
    AU  - Smith Stella Ifeanyi
    AU  - Fowora Adenike
    AU  - Nwosuh Chika Ignatius
    Y1  - 2014/10/30
    PY  - 2014
    N1  - https://doi.org/10.11648/j.avs.20140206.12
    DO  - 10.11648/j.avs.20140206.12
    T2  - Animal and Veterinary Sciences
    JF  - Animal and Veterinary Sciences
    JO  - Animal and Veterinary Sciences
    SP  - 169
    EP  - 174
    PB  - Science Publishing Group
    SN  - 2328-5850
    UR  - https://doi.org/10.11648/j.avs.20140206.12
    AB  - Aims: To determine the presence of plasmids in Listeria monocytogenes strains isolated from fresh raw milk from healthy cattle in Nigeria and also determine its role in antibiotics resistance since this organism is a food borne pathogen of public health importance. Methods and results: Isolation of Listeria monocytogenes was done using selective enrichment and isolation protocol, as recommended by United States Department of Agriculture (USDA) and Association of Analytical Chemists/ International Dairy Federation (AOAC/ IDF). Identification was by phenotypic characterization and confirmed using β-haemolytic activity, phosphatidyl inositol phospholipase-C (PIPLC) assay and polymerase chain reaction (PCR) . Strains identified as Listeria monocytogenes from the six zones were determined and results expressed as rate of prevalence in graph. Twenty-three strains of Listeria monocytogenes isolated from 305 samples of fresh raw milk were screened for the presence of plasmid DNA using the method of Ehrenfeld and Clewel and agarose gel electrophoresis. One isolate was found to harbour one plasmid with molecular weight of 14.62kb. The plasmid was cured using different concentrations of acridine orange. Antibiotic sensitivity test was carried out on both the wild strain (with plasmid) and cured strain using disc diffusion method of Bauer-Kirby and it was found that the wild strain was resistant to seven of the twelve antibiotics used while the cured strain was resistant to only one. Conclusions: There is a possibility of the presence of strains of Listeria monocytogenes that can harbour plasmids. This could confer antibiotic resistance to the strains and thus make antibiotic therapy unsuccessful. The increase in the number of antibiotics which the cured strain of Listeria monocytogenes was susceptible to in this study showed that plasmid may play a significant role in antibiotics resistance. The Fulani herdsmen who take fresh raw milk without boiling or pasteurization may serve as carriers of resistant strains of Listeria monocytogenes and help in its spread in the environment through faecal pollution.
    VL  - 2
    IS  - 6
    ER  - 

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Author Information
  • National Veterinary Research Institute, Vom, Plateau State Nigeria

  • Department of Microbiology, Faculty of Science, University of Lagos, Lagos, Nigeria

  • Department of Microbiology, Faculty of Science, University of Lagos, Lagos, Nigeria

  • Molecular Biology and Biotechnology Division, Nigerian Institute for Medical Research, Yaba, Lagos, Nigeria

  • Molecular Biology and Biotechnology Division, Nigerian Institute for Medical Research, Yaba, Lagos, Nigeria

  • National Veterinary Research Institute, Vom, Plateau State Nigeria

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