Isolation and Antimicrobial Susceptability of Listeria Monocytogenes from Raw Milk and Milk Products in Northern Kaduna State, Nigeria

U. B. USMAN^1, , J. K. P. KWAGA², J. KABIR² and O. S. OLONITOLA³

¹Division of Veterinary Services, Ministry of Animal Health and Fisheries Development, Sokoto, Nigeria

²Department of Veterinary Public Health and Preventive Medicine, Faculty of veterinary medicine, Ahmadu Bello University, Zaria, Nigeria

³Department of Microbiology, faculty of Science, Ahmadu Bello University, Zaria, Nigeria

Cite this paper:
U. B. USMAN, J. K. P. KWAGA, J. KABIR and O. S. OLONITOLA. Isolation and Antimicrobial Susceptability of Listeria Monocytogenes from Raw Milk and Milk Products in Northern Kaduna State, Nigeria. Journal of Applied & Environmental Microbiology. 2016; 4(3):46-54. doi: 10.12691/jaem-4-3-1

Abstract

Listeria (L.) monocytogenes is an ‘emerging food borne pathogen’ that poses threat to global food safety. Its occurrence in milk and milk products constitute a negative impact to the dairy industry. Despite its veterinary public health significance, only few studies are available on the prevalence and characteristics of the organism from milk and milk products in Nigeria. The main objectives of this study were to isolate, identify and determine the antibiotic susceptibility patterns of the Listeria monocytogenes isolated from raw milk and milk product in Kaduna state Nigeria. A total of 550 samples of raw milk and milk products (fermented milk “Kindrimo” and milk butter “Manshanu”) were collected during a cross-sectional study. Of these 550 samples, 193 (35.10%) samples were positive for Listeria species- like growth when plated on chromogenic Listeria agar, 91 (47.15%) from raw milk, 65 (33.67%) from ‘Manshanu’ and 37 (19.15%) ‘Kindrimo’. A total of 36 (6.55%) were found to be L. monocytogenes based on conventional biochemical tests. However, when further subjected to Microbact^TM Listeria 12L detection Kit (Oxoid, MB1128A) only 3 (8.3%) of the 36 isolates were identified as L. monocytogenes, while the others were L. grayi 4 (11.1%), L. ivanovii 27 (75.0%) and L. seeligeri 2 (5.6%). The multiplex PCR assay identified 9 Listeria (L.) monocytogenes isolates harbouring the hly A gene. The susceptibility of 36 L. monocytogenes isolates to 10 antibiotics was evaluated using the disk diffusion method. Isolates from milk samples had an overall resistance of 64.09% to the antibiotics, followed by isolates from ‘Kindrimo’ (61.67%), and least resistance was observed in isolates from ‘Manshanu” (58.33%). Overall, L. monocytogenes isolates showed the highest frequency of resistance to ampicillin (100%), followed by penicillin (95%) and cloxacillin (90%). In conclusion, the isolation of L. monocytogenes and other Listeria species in this study calls for an improved hygienic practices in the milk and milk products production channels and for the enlightment of the Fulani herds men and women by agricultural extension workers. Also, the resistance pattern shown by the isolates is an indication that the use of antibiotics should be regulated to minimize the incidence of antibiotic resistance.

Keywords:
listeria monocytogenes isolation antibiotic resistance

This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

Figures

References:

[1]	Babatunde, G. M., Oyenuga, V. A. and Loosli, J. K. (1999). Problems and possibility for milk production. Animal Production in the Tropics, 1: 43-60.
[2]	Donkor, E. S., Aning, K. G. and Quaye, J. (2007). Bacterial contamination of informally marketed raw milk in Ghana. Ghana Journal 41: 58-61.
[3]	Scuk, 2007. Evaluation of community resettlement and reintegration Programme activities. Scuk Jigiga Area Programme. Addis Ababa, SC, UK Ethiopia.
[4]	Adetunji, V. O. and Arigbede, M. I. (2011). Occurrence of E. coli O157:H7 and Listeria monocytogenes and Identification of Hazard Analysis Critical Control Points (HACCPs) in Production Operations of a Typical Tropic Cheese ‘Wara’ and Yoghurt. Pakistan Journal of Nutrition, 10: 796-804
[5]	Savadogo, A., Ouattaracherk, A. T., Paul, W., Baro, N. O., S. and Aboubacar, T., Alfred, S. (2004). Microorganisms involved in Fulani fermented milk in Burkina Faso. Pakistani Journal of Nutrition, 3:134-139.
[6]	Graves, L. M., Heelsel, L. O., Steigerwait, A. G., Morey, R. E., Daneshvar, M. L., Roof, S. E., Orsi, R. H., Fortes, E. D. and Milillo, S.R. (2010). Listeria marthii spp. nov., isolated from the natural environment of Finger Lakes National Forest. International Journal of Systemic and Evolutionary Microbiology, 60: 1280-1288.
[7]	Leclercq, A., Clermont, D., Bizet, C., Drimont, B., Le Fleche-Mateos, A., Roche, S., Buchrieser, C., Cadet-Daniel, V., Le Monnier, A., Lecuit, M., and allerberger, F. (2009). Cited by Henk, C. D., Bundrant, B. N., Fortes, E. D., Orsi, R. H., and Wiedmann, M. (2010). A population genetics- based and phylogenetic approach to understanding the evolution of virulence in the genus Listeria. Applied and Environmental Microbiology, 76: 6085-6100.
[8]	Bertsch, D., Rau, J., Eugster, M. R., Haug, M. C., Lason, P. A., Lacroix, C. and Meile, I. (2013). Listeria fleischmannii spp nov., isolated from cheese. International Journal Systemic and Evolutionary Microbiology, 63: 526-532.
[9]	Lang Halter, E., Neuhanus, K. and Scherer, S. (2013). Listeria weihenstephanensis sp novi., isolated from the water plant Lemma trisulca taken from a freshwater pond. International Journal of Systemic and Evolutionary Microbiology, 63: 641-647.
[10]	Henk, C. B., Warchocki, S., Wright, E. M., Allied, A. F., Ahlstrom, C., Manuel, C. S., Stasiewicz, M. J., Burrel, A., Roof, S., Strawn, L. K., Fortes, E., Nightingale, K. K., Kephart, D. and Wiedmann, M. (2014). Five new species of Listeria (L. floridensis sp, nov, L. aquqtica sp, nov., L. cornellensis sp. Nov. L. riparia sp, nov. and L. grandensis sp. Nov.) from agriculture and natural environment in the United States. IJSEM Papers in Press, Published March 5, 2014.
[11]	Schmid, M. W., Ng, E. Y., Lapidis, R., Emmerth, M., Walcher, M., Kreft, J., Goebel, W., Wagner, M. and Schleifer, K. H. (2005). Evolutionary history of genus Listeria and its virulence genes. Systemic Applied Microbiology, 28; 1-18.
[12]	Dominquez-Bernal, G., Stefanie, M. A., Bruno G. Z., Mariela S., Petra, H., Hector, J. M., Lizeth, L. Jurgn K. And Vazque- Boland J. A. (2006). A spontaneous genomic deletion in Listeria ivanovii identities LIPI-2, a specie-specific pathogenicity island encoding sphingongelinase and numerous internalins. Molecular Microbiology, 59: 415-432.
[13]	Food and Agriculture Organisations (1999). Report of the FAO expert consultation on the trade Impact of Listeria in fish products. FAO fisheries report NO. 64Fillu/ESNS/R604.Amsherst, M.A United states PP 17-20
[14]	Abay, S., Aydin, F., and Sumerkan, A. B (2012). Molecular typing of Listeria spp. isolated from different sources. Ankara Üniv Vet FakDerg, 59: 183-190.
[15]	Olive,r S. P., Jayaro, B. M. and Almeida, R. A. (2005). Food-borne pathogens in milk and the dairy farm environment: food safety and public health implications. Foodborne Pathologic Disease, 2: 115.
[16]	Aygun O, and Serda, P. (2006). Listeria spp. in the raw milk and dairy products in Antakya, Turkey. Food Control, 17: 676-678.
[17]	Scallan, E., Hoekstra, R. M., Angulo, F. J., Tauxe, R. V., Widdowso, M. A., Roy, S. L., Jones, J. L., and Griffin P. M. (2011). Food- borne illnesses acquired in the United States-Major pathogens. Emerging Infectious Diseases, 17: 7–15.
[18]	Obi, C. N. and Ikenebom, M. J. (2007). Studies on the Microbiology and Nutritional Qualities of a Nigerian fermented milk product (Nono). International Journal of Dairy Science, 2, 95-99.
[19]	Cartwright, E., Jackson, K. A., Johnson, S. D., Graves, L. M., Silk, B.J. and Mahon. E. B. (2013). Listeriosis outbreaks and associated food vehicles, United States, 1998–. 2008. Emerging Infectious Disease, 19: 1-9.
[20]	Silk, B. J., Date, K. A., Jackson, K. A., Pouillot, R., Kristin, G. H., Lewis, M. G. (2012). Invasive listeriosis in Foodborne Diseases Active Surveillance Network (Food Net), 2004-2009: Further Targeted Prevention Needed for Higher-Risk Groups. Clinical Infectious Disease, 54: 396-404.
[21]	Jalali, M. and Abedi, D. (2008). Prevalence of Listeria species in food products in Isfahan, Iran. International Journal of Food Microbiology, 122: 336-340.
[22]	CDC (2013). Incidence and trends of infection with pathogens transmitted commonly through food. Foodborne Diseases Active Surveillance Network, 10 U.S. sites, 1996–2012. MMWR 62, 283-287.
[23]	Hof, H., Nichterlein, T. and Kretschmar, M. (1997). Management of listeriosis. Clinical Microbiology Review, 10: 345-357.
[24]	Morvan, A., Moubareck, C., Leclercq, A., Hervé-Bazin, M., Bremont, S., Lecuit, M. (2010). Antimicrobial resistance of Listeria monocytogenes strains isolated from humans in France. Antimicrobial Agents and Chemotherapy, 54; 272-2731.
[25]	Troxler,R.,vonGraevenitz, A., Funke,G.,Wiedemann, B., Stock, I. (2000). Natural antibiotic susceptibility of Listeria species: L. grayi, L. innocua, L. ivanovii, L. monocytogenes. seeligeri and L. welshimeri strains. Clinical. Microbiology Infection. 6, 525e535.
[26]	Pesavento, G. Doci, B. Nieri, O. Comodo, N. and Lo Nostro, A. (2010). Prevalence and antibiotic susceptibility of Listeria spp. isolated from raw meat and retail foods. Food Control. 21:708-713.
[27]	Roberts, D., W. Hoper and Greenwood, M. (1995). Practical food microbiology. London: Public health Laboratory Services, pp: 146-149.
[28]	Janzten, M. M., Navas, J., Corujo, A., Moreno, R., López, V. and Martínez-Suárez, J. V. (2006). Review. Specific detection of Listeria monocytogenes in foods using commercial methods: from chromogenic media to real-time PCR. Spanish Journal of Agriculture. Research, 4:235-247.
[29]	Border, P., M., Howard, J., J., Plastow, G., S. and Siggens, K., W. (1990). Detection of Listeria species and Listeria monocytogenes using polymerase chain reaction. Letters in Applied Microbiology; 11:158-162.
[30]	Bauer, A., Kirby, W., Sherris, J., C., and Turck, M. (1966). Antibiotic susceptibility testing by a standardized single disk method. American journal of clinical pathology, 45(4):493.
[31]	(CLSI) Clinical Laboratory Standards Institute (2011). Performance standards for antimicrobial disk susceptibility tests; Approved standard- 9th Ed. CLSI document M2-A9. 26:1. Clinical Laboratory Standards Institute, Wayne, PA.
[32]	Conter, M., Paludi, D., Zanardi, E., Ghidini, S., Vergara, A. and Ianieri, A. (2009). Characterization of antimicrobial resistance of foodborne Listeria monocytogenes. International Journal of Food Microbiology, 128: 497-500.
[33]	Singh, S., Vadar, Singh, S. and Bharti, P. (2010). Prevalence of Salmonella in chicken eggs collected from poultry farms and marketing channels and their antimicrobial resistance. Food Research International, 43:2027-2030.
[34]	Jamali, H., Chai, L. C. and Thong, K. L. (2013). Detection and isolation of Listeria spp. and Listeria monocytogenes in ready-to-eat foods with various selective culture media. Food Control, 32: 19-24.
[35]	Boubendir, A., Hamidechi, M. A., Mostakim, M., El Abed, S., Ibnsouda Koraichi, S. (2011). Incidence of Listeria species and other psychrotrohic bacteria in bovine raw milk products in Northern Eastern Algeria. Medical Veterinary Review, 162:265-269.
[36]	Ning, N., Guo, K., Chug, L., Xu, L., Zhag, C., Cui, H., Cheng, Y., R., Liu, Q., Cio, W. and Zheng, Y. (2013). Pilot survey of raw whole milk in China from L. monocytogenes using PCR. Food Control, 31:176-179.
[37]	Benkeroum, N., Oubel, H., Zahar, M., Dlia, S. and Filali- Maltoul, A. (2003). Isolation of a bacteriocin producing Lactococcus lactis subsp. Lactis and application to control Listeria monocytogenes in Moroccan jben. Journal of Applied Microbiology, 89 (6): 960-968.
[38]	Adetunji, V. O. and Olaoye, O. O. (2012). Incidence and antibiotic susceptibility of Listeria monocytogenes isolates from milk of West African dwarf and red Sokoto breeds of goat from South-western Nigeria. New York Science Journal, 5 (11): 68-73.
[39]	Yakubu, Y., Salihu, M. D., Faleke, O. O., Abubakar, M. B., Junaidu, A. U., Magaji, A. A., Gulumbe, M. L. and Aliyu, R. M. (2012). Prevalence and antibiotic susceptibility of Listeria monocytogenes in raw milk from cattle herds within Sokoto Metropolis, Nigeria. Sokoto Journal of Veterinary Sciences, 10:13-17.
[40]	Issa, Z. M., Mustakim, M., Mohamed, S. A. S., Muda, N. M., Yen, L. H. and Radu, S. (2011). Antibiograms profiles of Listeria monocytogenes isolated from foods. International Conference on Biotechnology and Food Science (IPCBEE), 7: 133-137.
[41]	Kalorey, D. R., Warke, S. R., Kurkure, N. V., Rawool, D. B. and Barbuddhe, S. B. (2008). Listeria species in bovine raw milk: A large survey of Central India. Food Control, 19: 109-112.
[42]	Sauders, B. D., Durak, M. Z., Fortes, E., Windham, K., Schukken, Y. and Lembo, A. J. (2006). Molecular characterisation of Listeria monocytogenes from natura l and urban environment. Journal of Food Protection, 69: 93-105.
[43]	Kasalica, A., Vukovic, V., Vranjes, A. and Memisi, N. (2011). Listeria monocytogenes in milk and dairy products. Biotechnology in Animal Husbandry, 27: 1067-1082.
[44]	Ndahi, M. D., Kwaga, J. K. P., Bello, M., Kabir, J., Umoh, V. J., Yakubu, S. E. and Nok A. J. (2013). Prevalence and antimicrobial susceptibility of Listeria monocytogenes and methicillin-resistant Staphylococcus aureus strains from raw meat and meat products in Zaria, Nigeria. Letters in Applied Microbiology, 58(3): 263-269.
[45]	Al sheikh, A. D. I., Mohammed, G. E. and Abdalla, A. (2012). Isolation and identification of L. monocytogenes from fresh raw dressed broiler chicken in Sudan. Research Journal of Microbiology, 7: 319-326.
[46]	Perrin, M., Berner, M. and Delamare, C. (2003). Fatel case of Listeria innocua bacteraemia. Journal of Clinical Microbiology, 41: 5308-5309.
[47]	Gasanov, U., Hughes, D. and Hansbro, M. P. (2005). Methods for the isolation and identification of Listeria spp. and Listeria monocytogenes: a review. FEMS Microbiology Reviews, 29: 851-875.
[48]	Christelle, G., Join-Lambert, O., Monnier, A., Leclercq, A., Mechai, F., Mamzer- Bruneel, M., Bielecka, M., Scortti, Mariela, Dission, O. Berche, P., Vazquez- Boland, J., Lortholary, O. and Lecuit, M. (2010). Human listeriosis caused by Listeria ivanovii. Emerging infectious Diseases, 166(1): 136-138.
[49]	Jay, J. (1996). Modern food microbiology. In: Heldman DR, editor. Foodborne Listeriosis. New York: Chapman& Hall, pp: 478-506.
[50]	Mengaud, J., M. F. Vicente, J. Chenevert, J. Pereira, C. Geoffroy, B. Gicquel-Sanzey, F. Baquero, J. C. Perez-Diaz and and P. Cossart, (1988). Expression in Escherichia coli and sequence analysis of the listeriolysin O determinant of Listeria monocytogenes. Infection and Immunity, 56: 766-772.
[51]	Kuan, C., H., Goh, S., G., Loo, Y., Y., et al. (2013). Prevalence and quantification of L. monocytogenes in chicken offal at retail level in Malaysia. Poultry Science 2013; 92: 1664-1669.
[52]	Jallewor, P., K., Kalorey, D., R., Kurkure, N., V., Pande, V., V. and Barbuddhe, S., B. (2007). Genotypic characterisation of Listeria spp isolated from fresh water fish. International Journal of Food Microbiology, 114: 120-123.
[53]	Cooray, K. J., Nishibori, T., Xiong, H., Matsuyama, T., Fujita, M., and Mitsuyama, M. (1994). Detection of multiple virulence associated genes of Listeria monocytogenes by PCR in artificially contaminated milk samples. Applied and Environmental Microbiology, 60: 3023–3026.
[54]	Ieren, I. I., Bello, M. and Kwaga, J. K. P. (2013). Occurrence and antibiotic resistance profile of Listeria monocytogenes in salad vegetables and vegetable salads sold in Zaria, Nigeria. African Journal of Food Science, 7: 334-338.
[55]	Morobe, I. C., Obi, C. I., Nyila, M. A., Mataheka, M. I. and Gashe, B. A. (2009). Prevalence and antimicrobial resistance profiles of L. monocytogenes from various foods in Gaborone, Botswana. African Journal of Biotechnology, 8: (22)6383-6387.
[56]	Aureli, P., Ferrini, A. M. and Mannoni, V. (2003). Susceptibility of L. monocytogenes isolated from food in Italy to antimicrobial agents. International Journal of Food Microbiology, 83: 325-330.
[57]	Ennaji, H. H., Timinouni, M., Ennaji, M., Hassar, M. and Cohen, N. (2008). Characterization and antibiotic susceptibility of Listeria monocytogenes isolated from poultry and red meat in Morocco. Infection and Drug Resistance, 1: 45-50. 26.
[58]	Charpentier, E., Gerbaud, G., Jacquect, C., Rocourt, J. and Courvalin, P. (1995). Incidence of antibiotic resistance in Listeria spp. Journal of Infectious Diseases, 172: 277-289.
[59]	Levy, S. B. (2014). Reduced antibiotics use in livestock: How Denmark tackled resistance. Environmental Health Perspectives, 122. 6.
[60]	Adzitey, F., Hudu, N. and Ali, G. R. R. (2012). Prevalence and antibiotic resistance of Campylobacter, Salmonella and Listeria monocytogenes in ducks- A review. Foodborne Pathogen Disease, 9: 498-505.
[61]	Poyart-Salmeron, C., Carlier, C., Trieu-Cuot, P. and Courvalin, P. (1990). Transferable plasmid-mediated antibiotic resistance in Listeria monocytogenes. Lancet, 335: 1422-1426.
[62]	Wong, W. C., Pui, C. F., Tunung, R., Ubong, A., Noor Hidayah, M. S., Farinazleen, M. G., Noorlis, A., Cheah, Y. K. and Son, R. (2012). Antibiograms pattern among cultures of Listeria monocytogenes isolated from frozen burger patties in Malaysia. Pertanika. Journal of Tropical Agricultural Science, 35 (4): 793 – 804.
[63]	Krumperman, P. H. (1983). Multiple antibiotics resistance indexing Escherichia coli to identify risk sources of faecal contamination of foods. Applied Environmental Microbiology, 46: 165-170.
[64]	Safdar. A. and Arms Armstrong, D. (2003). Antimicrobial activities against 84 Listeria monocytogenes isolates from patient with systemic listeriosis at a comprehensive cancer centre (1955-1997). Journal of clinical microbiology, 41:483-85
[65]	Adzitey, F., Huda, Ali, N. G. R. R. and Manual, C. S. (2013). Isolation and characterisation of unidentified Listeria strains showing phenotypic and genetic similarities to a novel group of Listeria species from Malaysian Peking ducks. Journal of Biological Sciences, 13: 614-620.