Open Access Open Access  Restricted Access Subscription Access
Open Access Open Access Open Access  Restricted Access Restricted Access Subscription Access

Isolation and Identification of mecA gene from MRSA isolated from Local Sheep and Evaluate the Inhibitory effect of Cranberry Leaves extract In-Vitro


Affiliations
1 Department of Physiology and Pharmacology, College of Veterinary Medicine, Wasit University, Wasit, Iraq
2 Department of Microbiology, College of Veterinary Medicine, Wasit University, Wasit, Iraq
     

   Subscribe/Renew Journal


The current study focused and gave an insight about the distribution percentage of Methicillin-Resistant Staphylococcus aureus or sometimes called (MRSA) in sheep admitted to Veterinary medicine college, Wasit university at the period extended from March 2017 to June 2018 in Wasit province, Iraq and evaluate the inhibitory activity of ethanol extract of cranberry leaves. Consequently, from a total one hundred and fifty specimens, seventy samples were positive for the presence of S. aureus. Moreover, higher rate was detected in the wounds samples 37 (74%) and respiratory samples 20(40%) higher than that found in urine 10 (20%). The results of antibiotics susceptibility test for Staphylococcus aureus were indicated a high and moderate level of resistance to all antibiotics used. Additionally, a polymerase chain reaction was used, as a molecular technique, to detect mecA and results indicated that 54 strains were mecA positive and were labelled as MRSA. Moreover, results indicated that cranberry leaves extract had inhibitory activity against MRSA. In conclusion, the leaves extract of Cranberry has shown powerful antimicrobial activity, and these finding might be encouraging to several applications as antimicrobials or animals feed preservation.

Keywords

MSRA, PCR, mecA, Bacterial Resistance, Cranberry Leaves.
Subscription Login to verify subscription
User
Notifications
Font Size


  • Enright MC, Robinson DA, Randle G, Feil EJ, Grundmann HB, Spratt G. The evolutionary history of methicillin-resistant Staphylococcus aureus (MRSA). Proc. Natl. Acad. Sci. USA 2002; 99: 7687-7692.
  • Kuehnert MJ, Hill HA, Kupronis BA, Tokars JI, Solomon SL, Jernigan DB. Methicillin-resistant Staphylococcus aureus hospitalizations, United States. Emerg. Infect. Dis. 2005; 11: 868-872.
  • Weese JS, Caldwell F, Willey BM, Kreiswirth BN, McGeer A, Rousseau J. An outbreak of methicillin-resistant Staphylococcus aureus skin infections resulting from horse to human transmission in a veterinary hospital. Veterinary Microbiology 2006; 114: 160-164.
  • Griffeth GC, Morris DO. Screening for skin carriage of methicillin resistant coagulase-positive Staphylococci and Staphylococcus schleiferi in dogs with healthy and inflamed skin. Veterinary Dermatology 2008; 19: 142-l49.
  • Subhadharsini S, Gopinath P. Detection of the Presence of Bbp Gene for Bone Bound Sialoprotein among Clinical isolates of Staphylococcus aureus. Research J. Pharm. and Tech 2016; 9(9):1430-1432.
  • Mulligan ME, Murray-Leisure KA, Ribner BS, Standiford HC, John JF, Korvick JA, Kauffman CA, Yu VL. Methicillin-resistant Staphylococcus aureus: a consensus review of the microbiology, pathogenesis, and epidemiology with implications for prevention and management. Am. J. Med. 1993; 94: 313-328.
  • Pinho MG, Filipe SR, Lencastre H De, Tomasz A. (): Complementation of the essential peptidoglycan transpeptidase function of penicillin-binding protein 2 (PBP2) by the drug resistance protein PBP2A in Staphylococcus aureus. J. Bacteriol. 2001; 183-6525-6531.
  • Lowy FD. Antimicrobial resistance: the example of Staphylococcus aureus. J. Clin. Invest. 2003; 111: 1265-1273.
  • Rich M. Staphylococci in animals: prevalence, identification and antimicrobial susceptibility, with an emphasis on methicillin-resistant S. aureus. Brit. J. Biomed. Sci. 2005; 62: 98-105.
  • Broekema NM, Van TT, Monson TA, Marshall SA, Warshauer DM. Comparison of cefoxitin and oxacillin disk diffusion methods for detection of mecAmediated resistance in Staphylococcus aureus in a large-scale study. J. Clin. Microbiol. 2009; 47: 217-219.
  • Tiwari HK, Sapkota D, Das AK, Sen MR. Assessment of different tests to detect methicillin resistant Staphylococcus aureus. Southeast Asian J. Trop. Med. Publ. Health 2009; 40: 801-806.
  • F. Mariyam Niyas, Dr. Gopinath P. Comparative study on detection of MRSA using oxacillin agar screening method, cefoxitin disc diffusion method and mecAgene by PCR among clinical isolates of Staphylococcus aureus. Research J. Pharm. and Tech 2016; 9(9):1317-1320.
  • Choi SM, Kim S, Kim C, Lee D, Choi J, Yoo J, Kang J, Shin W, Kang W. Multiplex PCR for the detection of genes encoding aminoglycoside modifying enzymes and methicillin resistance among Staphylococcus species. J. Korean Med. Sci. 2003; 18: 631-636.
  • Kalhor H, Shariati L, Validi M, Tabatabaiefar MA, Nafisi MR. Comparison of agar screen and dublex-PCR methods in determination of methicillinresistant Staphylococcus aureus (MRSA) strains isolated from nasal carriage. African J. Microbiol. Res. 2012; 6: 3722-3726.
  • Goossens H. The Chennai declaration on antimicrobial resistance in India. Lancet Infect Dis 2013; 13:105–106.
  • Ayodele SQ. The effects of herbal remedies. Paper Presented at the 12th annual Conference of the Botannicial Socity of Nigeria (BOSON) University of Lago 2003.
  • Manjunath Sangappa, Padma Thiagarajan. Isolation and Screening of Soil Penicilliumsp VIT-2012 Metabolites against Methicillin Resistant Staphylococcus aureus. Research J. Pharm. and Tech. 6(12): Dec. 2013; Page 1340-1349.
  • Klein MA. Cranberry (Vacciniummacrocarpon) Aiton. In: Encyclopedia of Dietary Supplements; Coates, P., Blackman, M.R., Cragg, G., Levine, M., Moss, J., White, J., Eds.; Marcel Dekker: New York, Pp. 2005; 143- 149.
  • Jepson RG, Craig JC. Cranberries for preventing urinary tract infections. Cochrane Database Syst. Rev. 2008; 23: 2-8.
  • Forbes BA, Sahm DF, Weissfeld AS. Bailey and Scott’s Diagnostic Microbiology, Mosby Inc, Maryland Heights, Mo, USA, 12th edition, 2007.
  • CLSI. Performance standards for antimicrobial susceptibility testing. M100, 28th ed. January 2018. Wayne, PA 19087 USA: Clinical and Laboratory Standards Institute. 2018.
  • Kaya EG, Karakos E, Yagci S, Yucel M. Evalution of phenotypic and genotypic methods for detection of methicillin resistance in Staphylococcus aureus. African j. of Microbiologyy Research. 2009; 3: 925-929.
  • Effraim D, Salami HA, Osewa, TS. The effect of aqueous leaf extract of Ocimumgratissimum on haematological and biochemical parameters in rabbits. Afr. J. Biomed. Res. 2000; 3: 175-179.
  • Sanjay R Patel, Ashok P Suthar, Anand M Shah, Hitesh V Hirpara, Vishal D Joshi , Mayur V Katheria. Antimicrobial Activity of Methanolic Extracts of Mucunapruriens, Semecarpusanacardium, Anethumgraveolens by Agar Disc Diffusion Method. Research J. Pharm. and Tech. 3(1): Jan. - Mar. 2010; Page 165-167.
  • Miller LG, Diep BA. Clinical practice: colonization, fomites, and virulence: rethinking the pathogenesis of community associated methicillin-resistant Staphylococcus aureus infection. Clin Infect Dis 2008; 46: 752–760.
  • Graveland H, Duim B, van Duijkeren, E, Heederik D, Wagenaar A. Livestock-associated methicillin-resistant Staphylococcus aureus in animals and humans. Int J Med Microbiol 2011; 301: 630–634.
  • Witte W, Strommenger B, Stanek C, Cuny C. Methicillin resistant Staphylococcus aureusST398 in humans and animals, Central Europe. Emerg. Infect. Dis. 2007; 13: 255–258 .
  • Golding GR, Bryden L, Levett PN, McDonald RR, Wong A, Wylie J, Graham MR, Tyler S, van Domselaar G, Simor AE, Gravel D, Mulvey MR. Livestock-associated methicillin-resistant Staphylococcus aureus sequence type 398 in humans, Canada. Emerg. Infect. Dis. 2010; 16: 587–594 .
  • Gómez-Sanz E, Simón C, Ortega C, Gómez P, Lozano C, Zarazaga M, Torres C. First detection of methicillin-resistant Staphylococcus aureus ST398 and Staphylococcus pseudintermedius ST68 from hospitalized equines in Spain. Zoonoses Public Health 2014; 61: 192–201 .
  • Khamael MA, Saad SH. Molecular detection of Staphylococci Isolated from Mastitis in Sheep and Cows in Thi-Qar province. IJRANSS. 2015; 3: 33-44.
  • Gulani IA, Yaqub AG, Lawan A, Lawal JR, Falmata AA. Prevalence and phenotypic detection of methicillin resistance Staphylococcus aureus between ruminants butchered for humanoid intake and animal handlers in Maiduguri, Nigeria. J. Adv. Vet. Anim. Res., 2016; 3: 152-159.
  • Deepa S, R. Bharathidasan, K.Kanimozhi, A. Panneerselvam. Bacteriological Profile of Surgical and Non Surgical Wound Infections in a Tertiary Care Hospital. Research J. Pharm. and Tech. 5(6): June 2012; Page 853-856.
  • Vane Swetah C.S., Gopinath Prakasam. Detection of etaB gene for the production of exfoliative toxin among clinical isolates of Staphylococcus aureus. Research J. Pharm. and Tech 2016; 9(9):1469-1471.
  • Khushali K. Shah, Gopinath P. Detection of FnbA gene encoding for fibronectin binding protein among clinical isolates of Staphylococcus aureus. Research J. Pharm. and Tech. 2017; 10(2): 378-380.
  • Mai-siyama IB, Okon KO, Adamu NB., Askira UM, Ishaka TM, Adamu SG, Mohammed A. Methicillin-resistant Staphylococcus aureus (MSRA) colonization rate among ruminants’ animals slaughtered for human consumption and contact persons in Maiduguri, Nigeria. African Journal of Microbiology Research, 2014; 8: 2643-2649.
  • Khudaier BY, Basil AA, Amaal MK. Detection of Methicillin Resistant Staphylococcus aureus Isolated from Human and Animals in Basrah Province / Iraq. MRVSA2013; 2: 12-21.
  • Hanon BM. Comparative study to the Staphylococcus aureus, isolated from the bovine and human with detection of virulence (coa) gene in these isolates by polymerase chain reaction. M.Sc, Basrah University, College of Veterinary Medicine, Basrah, Iraq, 2009 .
  • Krishnan PU, Miles K, Shetty N. Detection of methicillin and mupirocin resistance in Staphylococcus aureus isolates using conventional and molecular methods: a descriptive study from a burns unit with high prevalence of MRSA. J ClinPathol 2002; 55: 745–748.
  • Cauwelier B, Gordts B, Descheemaecher P, Van Landuyt H. Evaluation of disk diffusion method with cefoxitin (30 μg) for detection of methicillin resistant Staphylococcus aureus. Eur. J. Clin. Microbiol. Infect. Dis. 2004; 23: 389-392.
  • Jain A, Agarwal J, Bansal S. Prevalence of methicillin-resistant, coagulasenegative stap-hylococci in neonatal intensive care units: findings from a tertiary care hospital in India. J Med Microbiol, 2004; 53: 941-44.
  • Rallapalli S, Verghese S, Verma RS. Validation of multiplex PCR strategy for simultane-ous detection and identification of methicillin resistant Staphylococcus aureus. Indian J Med Micro, 2006; 26: 361-64.
  • Rahbar M, Yaghoobi M, Fattahi A. Comparison of different laboratory methods for detection of methicillin resistant Staphylococ-cusaureus. Pak J Med Sci, 2006; 22: 442-445.
  • Rahimi F, Bouzari M, Vandyousefi J, Maleki Z, SaberiKashani S, Davoudi S. Analysis of antibiotic resistance and detection of mecA gene in Staphylococcus aureus isolated from hospitals and medical laboratories in Tehran. Irn J Biol, 2009; 21: 64-73.
  • Cakir N. Rasy onelolmayan antibiyotik kullanımınınekonomiksonuçları. Klimik Dergisi 2001; 14: 35-40.
  • Hare KT, Das AK, Sapkota D, Sivarajan K, Pahwa VK. Methicillin resistant Staphylococcus aureus: prevalence and antibiogram in a tertiary care hospital in western Nepal.J. Infect. Dev. Ctries. 2009; 3: 681-684.
  • Samia P, Barakzi Q, Farooqi BJ, Nazia K, Sabir, N. Antimicrobial susceptibility pattern of clinical isolates of methicillin resistant Staphylococcus aureus. J. Pak. Med. Asso. 2007; 57:2-4.
  • Nathan C, Cars O. Antibiotic resistance – problems, progress, and prospects. N Engl J Med. 2014; 371: 1761–3.
  • Hatano T, Kusuda M, Inada K, Ogawa TO, Shiota S, Tsuchiya T, Yoshida T. Effects of tannins and related polyphenols on methicillin-resistant Staphylococcus aureus. Phytochemistry, 2005; 66:2047-2055.
  • Cushnie TPT, Lamb AJ. Recent advances in understanding the antibacterial properties of flavonoids. Int. J. Antimicrob. Agents, 2011; 38: 99-107.
  • Roccaro AS, Blanco AR, Giuliano F, Rusciano D, Enea V. Epigallocatechin-gallate enhances the activity of tetracycline in staphylococci by inhibiting its efflux from bacterial cells. Antimicrobial Agents and Chemotherapy, 2004; 48: 1968-1973.
  • Daglia M. Polyphenols as antimicrobial agents. Current Opinion in Biotechnology, 2012; 23: 174-181.
  • Fast, W, Sutton LD. Metallo-beta-lactamase: Inhibitors and reporter substrates. Biochimica Et Biophysica Acta-Proteins and Proteomics, 2013; 1834: 1648-1659.

Abstract Views: 246

PDF Views: 0




  • Isolation and Identification of mecA gene from MRSA isolated from Local Sheep and Evaluate the Inhibitory effect of Cranberry Leaves extract In-Vitro

Abstract Views: 246  |  PDF Views: 0

Authors

Sarhan R. Sarhan
Department of Physiology and Pharmacology, College of Veterinary Medicine, Wasit University, Wasit, Iraq
Hussein A. Mohammed
Department of Microbiology, College of Veterinary Medicine, Wasit University, Wasit, Iraq

Abstract


The current study focused and gave an insight about the distribution percentage of Methicillin-Resistant Staphylococcus aureus or sometimes called (MRSA) in sheep admitted to Veterinary medicine college, Wasit university at the period extended from March 2017 to June 2018 in Wasit province, Iraq and evaluate the inhibitory activity of ethanol extract of cranberry leaves. Consequently, from a total one hundred and fifty specimens, seventy samples were positive for the presence of S. aureus. Moreover, higher rate was detected in the wounds samples 37 (74%) and respiratory samples 20(40%) higher than that found in urine 10 (20%). The results of antibiotics susceptibility test for Staphylococcus aureus were indicated a high and moderate level of resistance to all antibiotics used. Additionally, a polymerase chain reaction was used, as a molecular technique, to detect mecA and results indicated that 54 strains were mecA positive and were labelled as MRSA. Moreover, results indicated that cranberry leaves extract had inhibitory activity against MRSA. In conclusion, the leaves extract of Cranberry has shown powerful antimicrobial activity, and these finding might be encouraging to several applications as antimicrobials or animals feed preservation.

Keywords


MSRA, PCR, mecA, Bacterial Resistance, Cranberry Leaves.

References