Emergence of Fluoroquinolone Resistance in Pigs, Nigeria

Authors: Igbakura I. Luga, Alex A. Adikwu

Abstract:

A comparison of resistance to quinolones was carried out on isolates of Shiga toxin-producing Escherichia coliO157:H7 from cattle and mecA and nuc genes harbouring Staphylococcus aureus from pigs. The isolates were separately tested in the first and current decades of the 21^st century. The objective was to demonstrate the dissemination of resistance to this frontline class of antibiotic by bacteria from food animals and bring to the limelight the spread of antibiotic resistance in Nigeria. A total of 10 isolates of the E. coli O157:H7 and 9 of mecA and nuc genes harbouring S. aureus were obtained following isolation, biochemical testing, and serological identification using the Remel Wellcolex E. coli O157:H7 test. Shiga toxin-production screening in the E. coli O157:H7 using the verotoxin E. coli reverse passive latex agglutination (VTEC-RPLA) test; and molecular identification of the mecA and nuc genes in S. aureus. Detection of the mecA and nuc genes were carried out using the protocol by the Danish Technical University (DTU) using the following primers mecA-1:5'-GGGATCATAGCGTCATTATTC-3', mecA-2: 5'-AACGATTGTGACACGATAGCC-3', nuc-1: 5'-TCAGCAAATGCATCACAAACAG-3', nuc-2: 5'-CGTAAATGCACTTGCTTCAGG-3' for the mecA and nuc genes, respectively. The nuc genes confirm the S. aureus isolates and the mecA genes as being methicillin-resistant and so pathogenic to man. The fluoroquinolones used in the antibiotic resistance testing were norfloxacin (10 µg) and ciprofloxacin (5 µg) in the E. coli O157:H7 isolates and ciprofloxacin (5 µg) in the S. aureus isolates. Susceptibility was tested using the disk diffusion method on Muller-Hinton agar. Fluoroquinolone resistance was not detected from isolates of E. coli O157:H7 from cattle. However, 44% (4/9) of the S. aureus were resistant to ciprofloxacin. Resistance of up to 44% in isolates of mecA and nuc genes harbouring S. aureus is a compelling evidence for the rapid spread of antibiotic resistance from bacteria in food animals from Nigeria. Ciprofloxacin is the drug of choice for the treatment of Typhoid fever, therefore widespread resistance to it in pathogenic bacteria is of great public health significance. The study concludes that antibiotic resistance in bacteria from food animals is on the increase in Nigeria. The National Food and Drug Administration and Control (NAFDAC) agency in Nigeria should implement the World Health Organization (WHO) global action plan on antimicrobial resistance. A good starting point can be coordinating the WHO, Office of International Epizootics (OIE), Food and Agricultural Organization (FAO) tripartite draft antimicrobial resistance monitoring and evaluation (M&E) framework in Nigeria.

Keywords: Fluoroquinolone, Nigeria, resistance, Staphylococcus aureus.

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1315549

References:

[1] World Health Organization. “WHO Advisory Group on Integrated Surveillance of Antimicrobial Resistance (AGISAR). Critically important antimicrobials for human medicine 3rd Revision 2011. WHO Document Production Services, Geneva, Switzerland, ”Clinical Infectious Disease, vol. 55, pp. 712–719, Oct. 2012
[2] R. J. Gosling, C. S. Clouting, L. P. Randall, R. A. Horton and R. H. Davies, “Ciprofloxacin resistance in E. coli isolated from turkeys in Great Britain,” Avian Pathology, vol. 41, pp. 83–89, Feb. 2012.
[3] I. Literak, M. Micudova, D. Tausova, A. Cizek, M. Dolejska, I. Papousek, J. Prochazka, J. Vojtech, F. Borleis, L. Guardone, S. Guenther, J. Hordowski, C. Lejas, W. Meissner, B. F. Marcos, and M. Tucakov, “Plasmid-mediated quinolone resistance genes in fecal bacteria from rooks commonly wintering throughout Europe, ”Microbial Drug Resistance, vol. 18, pp. 567–573, Nov. 2012.
[4] A. Agabou, N. Lezzar, Z. Ouchenane, S. Khemissi, D. Satta, A. Sotto, J. P. Lavigne and A. Pantel, “Clonal relationship between human and avian ciprofloxacin-resistant Escherichia coli isolates in North-Eastern Algeria,” European Journal of Clinical Microbiology and Infectious Diseases, vol. 35, pp. 227–234, Feb. 2016.
[5] M. Röderova, D. Halova, I. Papousek, M. Dolejska, M. Masarikova, V. Hanulik, V. Pudova, P. Broz, M. Htoutou-Sedlakova, P. Sauer, J. Bardon, A. Cizek, M. Kolar and I. Literak, “Characteristics of Quinolone Resistance in Escherichia coli Isolates from Humans, Animals, and the Environment in the Czech Republic,” Frontiers in Microbiology, vol. 7, pp. 2147, Jan. 2017.
[6] A. S. Fahrion, L. Jamir, K. Richa, B. Sonuwara, V. Rutsa, S. Ao, V. P. Padmakumar, R. P.Deka and D. Grace, “Food Safety Hazards in the Pork chain in Nagaland, North East India: Implication for human health,” International Journal of Environmental Research and Public Health, vol. 11, pp. 403-417, Jan. 2014.
[7] M. H. Brown, C. O. Gill, J. Hollingsworth, I. R. Nickelson, S. Seward, J. J. Sheridan, T. Stevenson, J. L. Sumner, D. M. Theno, W. R. Usborne, and D. Zink, “The role of microbiological testing in systems for assuring the safety of beef,” International Journal of Food Microbiology, vol. 62, pp.7–1, Dec. 2000.
[8] J. R. Ruby, J. Zhu, and S. C. Ingham. “Using indicator bacteria and Salmonella spp. test results from three large-scale beef abattoirs over an 18-month period to evaluate intervention system efficacy and plan carcass testing for Salmonella spp,” Journal of Food Protection, vol. 70, pp. 2732-2740, Sep. 2007.
[9] C. Zweifel, D. Baltzerand R. Stephan, “Microbiological contamination of cattle and pig carcasses at five abattoirs determined by swab sampling in accordance with EU decision 2001/471/EC,” Meat Science, vol. 69, pp. 559-566, Mar. 2005.
[10] G. R. Golding, L. Bryden, P. N. Levett, R. R. McDonald and A. Wong, “Livestock-associated methicillin-resistant Staphylococcus aureus sequence type 398 in humans, Canada, ”Emerging Infectious Diseases, vol. 16, pp. 587–594, Apr. 2010.
[11] H. F. Chambers, and F. R. Deleo, “Waves of resistance: Staphylococcus aureus in the antibiotic era”, Nature Reviews Microbiology, vol. 7, no. 9, pp. 629-641, Sep. 2009.
[12] T. Zhao, M. P. Doyle, J. Shere, and L. Garber, “Prevalence of enterohemorrhagic Escherichia coli O157:H7 in a survey of diary herds”, Applied and Environmental Microbiology, vol. 61 no. 4, pp. 1290-1293. April 1995.
[13] P. Vos, G. Garrity, D. Jones, N. R. Krieg, W. Ludwig, F. A. Rainey, and W. Whitman, Bergey's Manual of Systematic Bacteriology; New York: Springer-Verlag, 2009, ch 4.
[14] B. G. Weckman, and B. W. Catlin, “Deoxyribonuclease activity of Mirococci from clinical sources” Journal of Bacteriology, vol. 73, no. 6, pp. 747-753, June 1957.
[15] P. Mugg, S. Seymour, and S. Clark, “Efficiency of the New Microgen® Staph ID for the Identification of Medically Important Staphylococcus species” United Kingdom: Microgen biproducts limited, U. K. 2003.
[16] Danish Technical University, Multiplex PCR for the detection of the mecA gene and identification of Staphylococcus aureus, From the community reference library of antimicrobial resistance, DTU, 2014.
[17] A. W. Bauer, W. M. Kirby, J. C. Sherris, and M. Truch, “Antibiotic susceptibility testing by a standardized single disk method” American Journal of Clinical Pathology, vol. 45, no. 4, pp. 493-496, April 1966.
[18] J. A. Coghlan, J. G. Collee, R. Cruickshank, J. P. Duguid, R. R. Gilles, J. C. Gould, D. M. Green, and N. J. Hayward, Medical Microbiology: The Practice of Medical Microbiology, 12th ed. Churchill Livingstone, Edinburgh: 1975 pp. 587-588.
[19] K. Haugum, J. Johansen, C. Gabrielsen, L. T. Brandal, K. Bergh, D. W. Ussery, F. Drabløs, J. E. Afset, “Comparative Genomics to Delineate Pathogenic Potential in Non-O157 Shiga Toxin-Producing Escherichia coli (STEC) from Patients with and without Haemolytic Uremic Syndrome (HUS) in Norway,” PLoS ONE, vol. 9(10), e111788.https://doi.org/10.1371/journal.pone.0111788, Oct. 2014.
[20] M. d. Zohorul Islam, A. Musekiwa, K. Islam, S. Ahmed, S. Chowdhury, A. Ahad, “Regional Variation in the Prevalence of E. coli O157 in Cattle: A Meta-Analysis and Meta-Regression,” PLoS ONE, vol 9(4), pp. e93299, Apr. 2014.
[21] H. C. Lewis, K. Mølbak, C. Reese, F. M. Aarestrup, M. Selchau, M. Sørum, and R. L. Skov, “Pigs as source of methicillin-resistant Staphylococcus aureus CC398 infections in humans, Denmark”, Emerging Infectious Diseases, vol. 14 no. 9, pp. 1383–1389, Sep. 2008.
[22] E. de Boer, J. T. Zwartkruis-Nahuis, B. Wit, X. W. Huijsdens, A. J. de Neeling, T. Bosch, R. A. van Oosterom, A. Vila, A. E. Heuvelink, “Prevalence of methicillin-resistant Staphylococcus aureus in meat”, International Journal of Food Microbiology, vol. 134, no. 1-2, pp. 52–56, Aug. 2009.
[23] A. U. Nnachi, F. E. Emele, C. O. Ukaegbu, M. V. Agah, O. E. Udu-Ibiam, O.S. Chukwu, and M. M. Agwu, “Prevalence of methicillin-resistant Staphylococcus aureus in raw meat and meat handlers in Onitsha, Nigeria”, European Journal of Preventive Medicine, vol. 2, no. 1, pp. 9-15, 2014.
[24] V. Velasco, J. S. Sherwood, P. P. Rojas-Garcia, and C. M. Logue, “Multiplex Real-Time PCR for detection of Staphylococcus aureus, mecA and Panton-Valentine Leukocidin (PVL) genes from selective enrichments from animals and retail meat”, PloS ONE, vol. 9, no. 5, pp. e97617. May 2014.
[25] R. Ali, K. Al-Achkar, A. Al-Mariri, and M. Safi, “Role of polymerase chain reaction in detection of antibiotic-resistant Staphylococcus aureus”, The Egyptian Journal of Medical Human Genetics, vol. 15, pp. 293-298, June 2014.
[26] A. Costa, I. Kay, and S. Palladino, “Rapid detection of mecA and nuc genes in Staphylococci by Real-Time multiplex PCR”, Diagnostic Microbiology and Infectious Diseases, vol. 51, no. 1, pp. 13-17, Jan. 2005.
[27] ECDC (European Centre for Disease Prevention and Control), EFSA (European Food Safety Authority) and EMA (European Medicines Agency), “ECDC/EFSA/EMA first joint report on the integrated analysis of the consumption of antimicrobial agents and occurrence of antimicrobial resistence in bacteria from humans and food-producing animals,” Stockhom/parma/London ECDC/EFSA/EMA. EFSA Journalvol.13, pp. 4006, Jan. 2015.