Molecular detection of some tetracycline-resistant genes in Staphylococcus aureus isolated from sheep milk in Al-Qadisiyah province of Iraq
Ghassan Khudhair Esmael 1 * , Noor Mahmood Majeed 2
1 Veterinary Medicine College, University of Al-Qadisiyah, IRAQ2 Dentistry College, University of Al-Qadisiyah, IRAQ
Our study aims to investigate some tetracycline resistance genes in isolates of Staphylococcus aureus that collected from ewes milk that collected randomly. Sixty Milk samples are taken from ewes randomly and submitted to culturing on blood agar then submitted to PCR methods for final confirming detection of Staphylococcus aureus by using specific primers designed for (Glpf) gene depending on NCBI and Primer3 then making sensitivity test for tetracycline then make PCR for detection of some tetracycline resistance genes. Our results showed positive samples for Staphylococcus aureus was 14/60 (23.3%) by using PCR. Besides, all Staphylococcus aureus isolates were submitted to the tetracycline sensitivity test (disk diffusion method (Kirby‐Bauer) by tetracycline. Moreover, showing the percentage of tetracycline resistance isolates was 8/14(57.1%), and tetracycline sensitive isolates were 6/14 (42.8 %). Also, our results showed the percentage of tetracycline resistance genes tet(L) and tet(k) were 9/14 (64.2%) and 7/14 (50%) respectively, and percentage of isolates that carry gene tet(L) and tet(K) together was 2/14 (14.2)% by using polymerase chain reaction technique. Our study concluded ewes milk that contaminated by Staphylococcus aureus at a high rate, and the non-response to treatment by tetracycline is occurred due to present tetracycline resistance genes such as tet(L) and tet(k), were causes treatment failure by tetracycline.
tetracycline resistance genes ewes Staphylococcus aureus PCR
Ahmadi M, Rohani SMR, Ayremlou N (2010) Detection of Staphylococcus aureus in milk by PCR. Comparative Clinical Pathology 19(1): 91-94.
Akanbi OE, Njom HA, Fri J, Otigbu AC, Clarke AM (2017) Antimicrobial Susceptibility of Staphylococcus aureus Isolated from Recreational Waters and Beach Sand in Eastern Cape Province of South Africa. International journal of environmental research and public health, 14(9): 1001.
Al- Anbagi NA, Kshash Q (2013) Daignosis of Staphylococcus aureus mastitis in bovine in Al-Najaf province by using Polymerases chain reaction (PCR). AL-Qadisiya Journal of Vet. Med. Sci. 12(2).
Almobarak ME, Mohammed Salih RR and Gibreel HH (2020) In vitro efficacy of Tylosin and Enrofloxacin in treatment of bovine mastitis causing bacteria in Omdurman locality. Online Journal of Animal and Feed Research, 10(2): 53-58. https://doi.org/10.36380/scil.2020.ojafr7
Andrés F, C‐A, Cervantes JL, Cantú‐Soto EU, Sánchez‐Machado DI, Campas‐Baypoli ON (2017) PCR Assay for Detection of Staphylococcus aureus in Fresh Lettuce (Lactuca sativa), Intech. Open.
Arabzadeh F, Aeini F, Keshavarzi F, Karami P (2018) Resistance to Tetracycline and Vancomycin of Staphylococcus aureus Isolates from Sanandaj Patients by Molecular Genotyping. Ann Clin Lab Res 6(4): 260.
Ardic N, Ozyurt M, Sareyyupoglu B (2005) Investigation of erythromycin and tetracycline resistance genes in methicillin-resistant staphylococci. Int J Antimicrob Agents, 26: 213-218.
Bertrand X, Huguenin Y, Talon D (2002) First report of a catalase-negative methicillin-resistant Staphalococcus aureus, Diagnostic Microbiology and Infectious Disease, 43(3): 245-246.
Bismuth R, Zilhao R, Sakamoto H, Guesdon JL, Courvalin P (1990) Gene heterogeneity for tetracycline resistance in Staphylococcus spp. Antimicrob Agents Chemother 34(8): 1611-1614.
Boucher HW, Corey GR (2008) Epidemiology of methicillin-resistant Staphylococcus aureus. Clin Infect Dis, 46: 344-349.
Chopra I, Roberts M (2001) Tetracycline antibiotics: mode of action, applications, molecular biology, and epidemiology of bacterial resistance. Microbiol Mol Biol Rev, 65(2): 232-260.
Chopra I, Roberts M (2001) Tetracycline antibiotics: mode of action, applications, molecular biology, and epidemiology of bacterial resistance. Microbiol. Mol. Biol. Rev. 65: 232-260.
Emaneini M, Bigverdi R, Kalantar D, Soroush S, Jabalameli F, Noorazar Khoshgnab B, Asadollahi P, Taherikalani M (2013) Distribution of genes encoding tetracycline resistance and aminoglycoside modifying enzymes in Staphylococcus aureus strains isolated from a burn center. Annals of burns and fire disasters, 26(2): 76-80.
Esposito S, Leone S, Petta E (2009) Treatment options for skin and soft tissue infections caused by meticillin-resistant Staphylococcus aureus: oral vs. parenteral; home vs. hospital. Int J Antimicrob Agents, 34: 30-35.
Gitau W, Masika M, Musyoki M (2018) Antimicrobial susceptibility pattern of Staphylococcus aureus isolates from clinical specimens at Kenyatta National Hospital. BMC Res Notes 11: 226.
Gordon RJ, Lowy FD (2008) Review Pathogenesis of methicillin-resistant Staphylococcus aureus infection. Clin Infect Dis. 1(46 Suppl 5): S350-9.
Hedayatianfard K, Akhlaghi M, Sharifiyazdi H (2014) Detection of tetracycline resistance genes in bacteria isolated from fish farms using polymerase chain reaction. Veterinary research forum: an international quarterly journal, 5(4): 269-275.
Huys G, D'Haene K, Van Eldere J, von Holy A, Swings J (2005) Molecular diversity and characterization of tetracycline-resistant Staphylococcus aureus isolate from a poultry processing plant. Appl Environ Microbiol 71(1): 574-579.
Inglis B, Matthews PR, Stewart PR (1988) The expression in Staphylococcus aureus of cloned DNA encoding methicillin resistance. J Gen Microbiol 134(6): 1465-1469.
Jones CH, Tuckman M, Howe AY, Orlowski M, Mullen S, Chan K, Bradford PA (2006) Diagnostic PCR analysis of the occurrence of methicillin and tetracycline resistance genes among Staphylococcus aureus isolates from phase 3 clinical trials of tigecycline for complicated skin and skin structure infections. Antimicrob Agents Chemother 50(2): 505-510.
Kareem SD, Olusegun AK, Samuel OO (2017) The Effect of Co2 Emission and Economic Growth on Energy Consumption in Sub Sahara Africa. International Journal of Sustainable Energy and Environmental Research, 6(1): 27-35.
Li X, Huang T, Xu K (2019) Molecular characteristics and virulence gene profiles of Staphylococcus aureus isolates in Hainan, China. BMC Infect Dis 19: 873.
Lim KT, Hanifah YA, Yusof M, Thong KL (2012) ermA, ermC, tetM and tetK are essential for erythromycin and tetracycline resistance among methicillin resistant Staphylococcus aureus strains isolated from a tertiary hospital in Malaysia. Indian J Med Microbiol 30(2): 203-207.
Mandell GL, Douglas RG, Bennet JE (2015) Principles and practice of infectious diseases.
McCallum N, Berger-Bachi B, Senn MM (2010) Regulation of antibiotic resistance in Staphylococcus aureus. Int J Med Microbiol, 300: 118-129.
Naimi HM, Rasekh H, Noori AZ (2017) Determination of antimicrobial susceptibility patterns in Staphylococcus aureus strains recovered from patients at two main health facilities in Kabul, Afghanistan. BMC Infect Dis 17: 737.
Nwankwo EO, Nasiru MS (2011) Antibiotic sensitivity pattern of Staphylococcus aureus from clinical isolates in a tertiary health institution in Kano, Northwestern Nigeria. The Pan African medical journal, 8: 4.
Radostitis OM, Gay CC, Hinchliff KW (2007) Veterinary medicine a Texet book of the diseases of cattle, horses, sheep, pigs and got 10 Edition London: W. Saunder Company Limited.
Rainard P, Riollet C (2006) Innate immunity of bovine mammary gland. Vet. Res., (37): 369-400.
Roberts MC (2005) Update on acquired tetracycline resistance genes. FEMS Microbiol Lett; 245(2): 195-203.
Robicsek A, Beaumont JL, Paule SM, Hacek DM, et al. (2008) Universal surveillance for methicillin-resistant Staphylococcus aureus in 3 affiliated hospitals. Ann Intern Med. 18; 148(6): 409-418.
Rodriguez-Avial I, Rodriguez-Avial C, Culebras E, Picazo JJ (2003) Distribution of tetracycline resistance genes tet(M), tet(O), tet(L) and tet(K) in blood isolates of viridans group streptococci harbouring erm(B) and mef(A) genes. Susceptibility to quinupristin/dalfopristin and linezolid, International Journal of Antimicrobial Agents 21(6): 536-541.
Taponen S, Salmikivi L, Simojoki H, Koskinen MT, Pyörälä S (2009) Real-time polymerase chain reaction-based identification of bacteria in milk samples from bovine clinical mastitis with no growth in conventional culturing in milk. J. Dairy Sci., (92): 2610-2617.
Tripathi AB, Tancowny Q, Truong-Tran S, Retsema R, et al. (2006) Detection of Staphylococcus Aureus by Real-Time PCR in Chronic Rhinosinusitis with Nasal Polyps. Journal of Allergy and Clinical Immunology, 117(2): S71.
Trzcinski K, Cooper BS, Hryniewicz W, Dowson CG (2000) Expression of resistance to tetracyclines in strains of methicillin-resistant Staphylococcus aureus. Journal of Antimicrobial Chemotherapy, 45(6): 763-770.
Velasco V, Sherwood JS, Rojas-García PP, Logue CM (2014) 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 9(5): e97617.
Reference: Esmael GK, Majeed NM. Molecular detection of some tetracycline-resistant genes in Staphylococcus aureus isolated from sheep milk in Al-Qadisiyah province of Iraq. Eurasia J Biosci. 2020;14(1), 1161-1166.
In-text citation: (Esmael & Majeed, 2020)
Reference: Esmael, G. K., & Majeed, N. M. (2020). Molecular detection of some tetracycline-resistant genes in Staphylococcus aureus isolated from sheep milk in Al-Qadisiyah province of Iraq. Eurasian Journal of Biosciences, 14(1), 1161-1166.