الگوی مقاومت آنتی‌بیوتیکی و حضور ژن‌های حدت اپرون ica (مولد بیوفیلم) در استافیلوکوکوس آرئوس‌های کواگولاز مثبت جداشده از موارد ورم پستان گاوی در استان‌آذربایجان‌شرقی
محورهای موضوعی :
آسیب شناسی درمانگاهی دامپزشکی
سعید صالحی
1
(دانش‌آموخته دکترای حرفه‌ای دامپزشکی، دانشکده دامپزشکی، واحد تبریز، دانشگاه آزاد اسلامی، تبریز، ایران.)
یونس انزابی
2
(استادیار گروه پاتوبیولوژی، دانشکده دامپزشکی، واحد تبریز، دانشگاه آزاد اسلامی، تبریز، ایران.)
امیرعلی کاوه
3
(استادیار گروه علوم درمانگاهی، دانشکده دامپزشکی، واحد تبریز، دانشگاه آزاد اسلامی، تبریز، ایران.)
تاریخ دریافت : 1400/05/14
تاریخ پذیرش : 1400/09/01
تاریخ انتشار : 1400/08/01
کلید واژه:
استافیلوکوکوس اورئوس,
ورم پستان گاو,
کواگولاز مثبت,
اپرون ica,
بیوفیلم,
چکیده مقاله :
ورم پستان های استافیلوکوکی از جمله بیماری های مهم در دامداری های صنعتی می باشند و استافیلوکوک هایی که قادر به تولید بیوفیلم ، میکرو آبسه و ماکرو آبسه هستند ، عامل ورم پستان بددرمان تلقی شده و سبب حذف گاو ها می شوند . توانایی تولید بیوفیلم توسط باکتری های مذکور بستگی به وجود ژن های حدت اپرون ica (icaA، icaB،icaC ، icaD و icaR) و همچنین برخی فاکتور های محیطی دارد. در تحقیق حاضر الگوی مقاومت آنتی بیوتیکی و نیز وجود ژن های حدت اپرون icaدر استافیلوکوکوس آرئوس های جداشده از موارد ورم پستان گاوی بررسی شد. با توجه به یافته های به دست آمده از آزمایشات میکروبیولوژیکی و simplex PCR مشخص گردید که در گاوداری های مورد آزمایش در استان آذربایجان شرقی ، ورم پستان استافیلوکوکی شیوع نسبتا بالائی داشته و در میان جدایه های مربوطه، درصد نسبتاً بالائی از آن ها دارای انواعی از ژن های حدت اپرون ica هستند که توانایی تولید بیوفیلم را کد می کنند. همچنین نتایج آزمایش سنجش حساسیت آنتی بیوتیکی انجام شده در مورد جدایه های مذکور نشان دهنده مقاومت بیش از پیش آن ها نسبت به اکثر آنتی بیوتیک های مورد آزمایش بود. با توجه به این که بالا بودن میزان حضور ژن های اپرون ica مخصوصاً ژن هایی چون icaA و icaD منجر به تولید بیوفیلم قوی تر و بیشتر شده و نیز مقاومت نسبت به آنتی بیوتیک های مختلف هم افزایش می یابد. لذا، نتایج مطالعه حاضر نشان دهنده پیش آگهی نامناسبی در خصوص موفقیت درمان آنتی بیوتیکی ورم پستان های استافیلوکوکی در دامداری های منطقه می باشد.
چکیده انگلیسی:
Staphylococcal mastitis is one of the most important diseases in industrial livestock and Staphylococci capable of producing biofilm, micro and macro abscesses, are considered to be the cause of malignant mastitis which result in culling of dairy cattle. The ability of these bacteria to produce biofilm depends on the presence of ica operon genes (icaA, icaB, icaC, icaD and icaR) and also some environmental factors. In the present study, the pattern of antibiotic resistance and the presence of operon ica virulence genes in Staphylococcus aureus isolated from bovine mastitis was investigated. According to the findings of microbiological and simplex PCR tests, it was determined that in the farms tested in East Azerbaijan province, Staphylococcal mastitis has a relatively high prevalence and among the relevant isolates, a relatively high percentage of them contain a variety of operon ica virulence genes that encode the ability to produce biofilm. Also, the results of antibiotic susceptibility testing performed on these isolates showed that they were more resistant to most of the antibiotics tested. Due to the high presence of ica operon genes, especially genes such as icaA and icaD, which result in production of stronger and excessive biofilm as well as increased resistance to various antibiotics, the results of the present study therefore indicate an unfavorable prognosis regarding the success of antibiotic treatment of staphylococcal mastitis in livestock farms in the region.
منابع و مأخذ:
Anzabi, Y. and Shaghaghi, S. (2015). In vitro evaluation of antibacterial properties of propolis alcoholic extract on bovine mastitis isolates. Veterinary Clinical Pathology, 9(34): 117-129. [In Persian]
Arciola, C.R., Baldassarri, L. and Montanaro, L. (2001). Pesence of icaA and icaD genes and slime production in a collection of staphylococcal strains from catheter associated infections. Journal of Clinical Microbiology, 39(1): 2151-2156.
Beaudeau, F., Ducrocq, V., Fourichon, C. and Seegers, H. (1995). Effect of disease on length of productive life of french Holstein dairy cows assessed by survival analysis. Journal of Dairy Science, 78(1): 103-117.
Bradley, A.J. (2002). Bovine mastitis: an evolving disease. The Veterinary Journal, 164(2): 116-128.
Brakstad, O.G., Aasbakk, K. and Maeland, J.A. (1992). Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene. Journal of Clinical Microbiology, 30(7): 1654-1660.
Casagrande Proietti, P., Stefanetti, V., Hyatt, D.R., Marenzoni, M.L., Capomaccio, S., Coletti M., et al. Phenotypic and genotypic characterization of canine pyoderma isolates of Staphylococcusintermedius for biofilm formation. Journal of Veterinary Medicine Sciences, 77(8): 945-51.
Ciftci, A., Findik A., Emek Onuk E. and Savasan S. (2009). Detection of methicillin resistance and slime factor production of staphylococcus aureus in bovine mastitis. Brazilian Journal of Microbiology, 40(1): 254-261.
Clinical and Laboratory Standards Institute (2017). Performance standards for antimicrobial susceptibility testing; M02-A12, M07-A10 and M11-A8. M100. 27th ed., Wayne, U.S.A: Bublished by CLSI, pp: 12-16, 162-165.
Cramiton, S.E., Gerke, C. and Gotz, F. (2001). In vitro methods to study staphylococcal biofilm formation. Methods in Enzymology, 336(1): 239-255.
Esnaashari, M., Shayegh, J. and Nasrollahi Omran, A. (2012). Prevalence of genes encoding the classical enterotoxins of Staphylococcus aureus isolated from buffalo milk in Tabriz area by multipex PCR. Journal of Food Hygiene, 2(6): 61-68.
Frebourg, N.B., Lefebvre, S., Baert, S. and Lemeland, J.F. (2000). PCR-Based assay for discrimination between invasive and contaminating Staphylococcus epidermidis strains. Journal of Clinical Microbiology, 38(2): 877-80.
Gad, G.F., El-Feky, M.A., El-Rehewy, M.S., Hassan, M.A., Abolella, H. and El-Baky, R.M. (2009). Detection of icaA, icaD genes and biofilm production by Staphylococcus aureus and Staphylococcus epidermidis isolated from urinary tract catheterized patients. The Journal of Infection in Developing Countries, 3(5): 342-351.
Holtenius, K., Waller, K.P., Essen-Gustavsson, B., Holtenius, P. and Sandgren, C.H. (2004). Metabolic parameters and blood leukocyte profiles in cows from herds with high or low mastitis incidence. The Veterinary Journal, 168(1): 65-73.
Iorio, N.L., Lopes, A.P., Schuenck, R.P., Barcellos, A.G., Olendzki, A.N., Lopez, G.L., et al. (2011). A combination of methods to evaluate biofilm production may help to determine the clinical relevance of Staphylococcus in blood cultures. Microbiology and Immunology. 55(1): 28-33.
Kateete, D.P., Kimani, C.N., Katabazi, F.A., Okeng, A., Okee, M.S., Nanteza, A., et al. (2010). Identification of Staphylococcus aureus: DNase and Mannitol salt agar improve the efficiency of the tube coagulase test. Annals of Clinical Microbiology and Antimicrobials, 9(1): 23-24.
Khoramian, B., Emaneini, M.E., Bolourchi, M., Eslampour, M.A., Niasari-Naslaji, A., Aligholi, M., et al. (2010). Evaluation of the biofilm-forming ability of Staphylococcus aureus isolates from bovine mastitis in Iran. Journal of Comprative Pathobiology, 6(4): 109-114.
Kiani-Nia, M., Hasani, A., Hasani, A., Sharifi, Y., Ahmadi, S.M. and Deghani, L. (2013). Investigation and Identification of the nuc, femB, mecA and aac (6′)/aph(2′′)-Ia genes in the Staphylococcus aureus isolated from Northwest Iran by Multiplex PCR method. Medical Journal of Tabriz University of Medical Sciences and Health Services, 35(1): 68-73. [In Persian]
Liduma, I., Tracevska, T., Bers, U. and Zilevica, A. (2012). Phenotypic and genetic analysis of biofilm formation by Staphylococcus epidermidis. Medicina (Kaunas, Lithuania), 48(6): 305-309.
Nourbakhsh, F. and Momtaz, H. (2016). Evaluation of Phenotypic and Genotypic biofilm formation in Staphylococcus aureus isolates, isolated from Hospital infections in Shahrekord, 2015. Arak Medical University Journal, 19(109): 69-79. [In Persian]
Markey, B.K., Leonard, F.C., Archambault, M., Cullinane, A. and Maguire, D. (2013). Clinical Veterinary Microbiology. 2ed., Dublin: Mosby-Elsevier Ltd, pp: 445-453, 735-755.
Mirzaee, M., Najar-Peerayeh, S. and Ghasemian, A.M. (2014). Detection of icaABCD genes and biofilm formation in clinical isolates of methicillin resistant Staphylococcus aureus. Iranian Journal of Pathology, 9(4): 257-262.
Mirzaee, M., Najar-Peerayeh, S., Behmanesh, M., Forouzandeh-Moghadam, M. and Ghasemian, A.M. (2014). Biofilm formation and presence of ica genes in Staphylococcus aureus isolated from Intensive Care Unit. Journal of Mazandaran University of Medical Sciences, 24(115): 43-51. [In Persian]
Mirzaei, H., Javadi, A., Farajli, M., Shah-Mohammadi, A.R., Monadi, A.R. and Barzegar, A. (2012). Prevalence of Staphylococcus aureus resistant to methicillin in traditional cheese and cream: a study in city of Tabriz, Iran. Journal of Veterinary Research, 67(1): 65-70. [In Persian]
Moori-Bakhtiari, N. and Moslemi, M. (2017). Phenotypic evaluation of biofilm producing ability in Methicillin resistant Staphylococcus aureus. Journal of Kashan University of Medical Sciences (Feyz), 20(6): 525-231. [In Persian]
Peymani, A., Farajnia, S., Nahaei, M.R., Sohrabi, N., Abbasi, L., Ansarin, K., et al. (2012). Prevalence of Class 1 Integron among Multidrug-Resistant Acinetobacter baumannii in Tabriz, Northwest of Iran. Polish Journal of Microbiology, 61(1): 57-60.
Phuektes, P., Mansell, P. and Browning, G. (2001). Multiplex polymerase chain reaction assay for simultaneous detection of Staphylococcus aureus and Streptococcal causes of bovine mastitis. Journal of Dairy Science, 84(5): 1140-1148.
Quinn, P.J., Markey, B.K., Leonard, F.C., FitzPatrick, E.S., Fanning, S. and Hartigan, P.J. (2011).Veterinary Microbiology and Microbial Disease. Second ed., Dublin: Wiley-Blackwell Ltd, pp: 837-849.
Shahkarami, F. and Rashki, S. (2016). Prevalence of ica operon related genes in Staphylococcus aureus and Staphylococcus epidermidis clinical isolates. Iran Journal of Medical Microbiology, 9(4): 16-23. [in Persian]
Solati, S.M., Tajbakhsh, E., Khamesipour, F. and Gugnani H.C. (2015). Prevalence of virulence genes of biofilm producing strains of Staphylococcus epidermidis isolated from clinical samples in Iran. AMB Express, 5(47): 1-5.
Stevens, N.T., Tharmabala, M., Dillane, T., Greene, C.M., O’Gara, J.P. and Humphreys, H. (2008). Biofilm and the role of the ica operon and aap in Staphylococcus epidermidis isolates causing neurosurgical meningitis. Clinical Microbiology and Infection, 14(7): 719-722.
Suzuki, T., Kawamura, Y., Uno, T., Ohashi, Y. and Ezaki, T. (2005). Prevalence of Staphylococcus epidermidis strains with biofilm-forming ability in isolates from conjunctiva and facial skin. American Journal of Ophthalmology,140(5): 844-850
Vasudevan, P., Mohan-Nair, M.K., Annamalai, T. and Venkitanarayanan, K.S. (2003). Phenotypic and genotypic characterization of bovine mastitis isolates of Staphylococcus aureus for biofilm formation.Veterinary Microbiology, 92(1): 179-85.
Zmantar, T., Kouidhi, B., Miladi, H., Mahdouani, K. and Bakhrouf, A. (2010). A microtiter plate assay for Staphylococcus aureus biofilm quantification at various pH levels and hydrogen peroxide supplementation. New Microbiology, 33(2): 137-145.
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Anzabi, Y. and Shaghaghi, S. (2015). In vitro evaluation of antibacterial properties of propolis alcoholic extract on bovine mastitis isolates. Veterinary Clinical Pathology, 9(34): 117-129. [In Persian]
Arciola, C.R., Baldassarri, L. and Montanaro, L. (2001). Pesence of icaA and icaD genes and slime production in a collection of staphylococcal strains from catheter associated infections. Journal of Clinical Microbiology, 39(1): 2151-2156.
Beaudeau, F., Ducrocq, V., Fourichon, C. and Seegers, H. (1995). Effect of disease on length of productive life of french Holstein dairy cows assessed by survival analysis. Journal of Dairy Science, 78(1): 103-117.
Bradley, A.J. (2002). Bovine mastitis: an evolving disease. The Veterinary Journal, 164(2): 116-128.
Brakstad, O.G., Aasbakk, K. and Maeland, J.A. (1992). Detection of Staphylococcus aureus by polymerase chain reaction amplification of the nuc gene. Journal of Clinical Microbiology, 30(7): 1654-1660.
Casagrande Proietti, P., Stefanetti, V., Hyatt, D.R., Marenzoni, M.L., Capomaccio, S., Coletti M., et al. Phenotypic and genotypic characterization of canine pyoderma isolates of Staphylococcusintermedius for biofilm formation. Journal of Veterinary Medicine Sciences, 77(8): 945-51.
Ciftci, A., Findik A., Emek Onuk E. and Savasan S. (2009). Detection of methicillin resistance and slime factor production of staphylococcus aureus in bovine mastitis. Brazilian Journal of Microbiology, 40(1): 254-261.
Clinical and Laboratory Standards Institute (2017). Performance standards for antimicrobial susceptibility testing; M02-A12, M07-A10 and M11-A8. M100. 27th ed., Wayne, U.S.A: Bublished by CLSI, pp: 12-16, 162-165.
Cramiton, S.E., Gerke, C. and Gotz, F. (2001). In vitro methods to study staphylococcal biofilm formation. Methods in Enzymology, 336(1): 239-255.
Esnaashari, M., Shayegh, J. and Nasrollahi Omran, A. (2012). Prevalence of genes encoding the classical enterotoxins of Staphylococcus aureus isolated from buffalo milk in Tabriz area by multipex PCR. Journal of Food Hygiene, 2(6): 61-68.
Frebourg, N.B., Lefebvre, S., Baert, S. and Lemeland, J.F. (2000). PCR-Based assay for discrimination between invasive and contaminating Staphylococcus epidermidis strains. Journal of Clinical Microbiology, 38(2): 877-80.
Gad, G.F., El-Feky, M.A., El-Rehewy, M.S., Hassan, M.A., Abolella, H. and El-Baky, R.M. (2009). Detection of icaA, icaD genes and biofilm production by Staphylococcus aureus and Staphylococcus epidermidis isolated from urinary tract catheterized patients. The Journal of Infection in Developing Countries, 3(5): 342-351.
Holtenius, K., Waller, K.P., Essen-Gustavsson, B., Holtenius, P. and Sandgren, C.H. (2004). Metabolic parameters and blood leukocyte profiles in cows from herds with high or low mastitis incidence. The Veterinary Journal, 168(1): 65-73.
Iorio, N.L., Lopes, A.P., Schuenck, R.P., Barcellos, A.G., Olendzki, A.N., Lopez, G.L., et al. (2011). A combination of methods to evaluate biofilm production may help to determine the clinical relevance of Staphylococcus in blood cultures. Microbiology and Immunology. 55(1): 28-33.
Kateete, D.P., Kimani, C.N., Katabazi, F.A., Okeng, A., Okee, M.S., Nanteza, A., et al. (2010). Identification of Staphylococcus aureus: DNase and Mannitol salt agar improve the efficiency of the tube coagulase test. Annals of Clinical Microbiology and Antimicrobials, 9(1): 23-24.
Khoramian, B., Emaneini, M.E., Bolourchi, M., Eslampour, M.A., Niasari-Naslaji, A., Aligholi, M., et al. (2010). Evaluation of the biofilm-forming ability of Staphylococcus aureus isolates from bovine mastitis in Iran. Journal of Comprative Pathobiology, 6(4): 109-114.
Kiani-Nia, M., Hasani, A., Hasani, A., Sharifi, Y., Ahmadi, S.M. and Deghani, L. (2013). Investigation and Identification of the nuc, femB, mecA and aac (6′)/aph(2′′)-Ia genes in the Staphylococcus aureus isolated from Northwest Iran by Multiplex PCR method. Medical Journal of Tabriz University of Medical Sciences and Health Services, 35(1): 68-73. [In Persian]
Liduma, I., Tracevska, T., Bers, U. and Zilevica, A. (2012). Phenotypic and genetic analysis of biofilm formation by Staphylococcus epidermidis. Medicina (Kaunas, Lithuania), 48(6): 305-309.
Nourbakhsh, F. and Momtaz, H. (2016). Evaluation of Phenotypic and Genotypic biofilm formation in Staphylococcus aureus isolates, isolated from Hospital infections in Shahrekord, 2015. Arak Medical University Journal, 19(109): 69-79. [In Persian]
Markey, B.K., Leonard, F.C., Archambault, M., Cullinane, A. and Maguire, D. (2013). Clinical Veterinary Microbiology. 2ed., Dublin: Mosby-Elsevier Ltd, pp: 445-453, 735-755.
Mirzaee, M., Najar-Peerayeh, S. and Ghasemian, A.M. (2014). Detection of icaABCD genes and biofilm formation in clinical isolates of methicillin resistant Staphylococcus aureus. Iranian Journal of Pathology, 9(4): 257-262.
Mirzaee, M., Najar-Peerayeh, S., Behmanesh, M., Forouzandeh-Moghadam, M. and Ghasemian, A.M. (2014). Biofilm formation and presence of ica genes in Staphylococcus aureus isolated from Intensive Care Unit. Journal of Mazandaran University of Medical Sciences, 24(115): 43-51. [In Persian]
Mirzaei, H., Javadi, A., Farajli, M., Shah-Mohammadi, A.R., Monadi, A.R. and Barzegar, A. (2012). Prevalence of Staphylococcus aureus resistant to methicillin in traditional cheese and cream: a study in city of Tabriz, Iran. Journal of Veterinary Research, 67(1): 65-70. [In Persian]
Moori-Bakhtiari, N. and Moslemi, M. (2017). Phenotypic evaluation of biofilm producing ability in Methicillin resistant Staphylococcus aureus. Journal of Kashan University of Medical Sciences (Feyz), 20(6): 525-231. [In Persian]
Peymani, A., Farajnia, S., Nahaei, M.R., Sohrabi, N., Abbasi, L., Ansarin, K., et al. (2012). Prevalence of Class 1 Integron among Multidrug-Resistant Acinetobacter baumannii in Tabriz, Northwest of Iran. Polish Journal of Microbiology, 61(1): 57-60.
Phuektes, P., Mansell, P. and Browning, G. (2001). Multiplex polymerase chain reaction assay for simultaneous detection of Staphylococcus aureus and Streptococcal causes of bovine mastitis. Journal of Dairy Science, 84(5): 1140-1148.
Quinn, P.J., Markey, B.K., Leonard, F.C., FitzPatrick, E.S., Fanning, S. and Hartigan, P.J. (2011).Veterinary Microbiology and Microbial Disease. Second ed., Dublin: Wiley-Blackwell Ltd, pp: 837-849.
Shahkarami, F. and Rashki, S. (2016). Prevalence of ica operon related genes in Staphylococcus aureus and Staphylococcus epidermidis clinical isolates. Iran Journal of Medical Microbiology, 9(4): 16-23. [in Persian]
Solati, S.M., Tajbakhsh, E., Khamesipour, F. and Gugnani H.C. (2015). Prevalence of virulence genes of biofilm producing strains of Staphylococcus epidermidis isolated from clinical samples in Iran. AMB Express, 5(47): 1-5.
Stevens, N.T., Tharmabala, M., Dillane, T., Greene, C.M., O’Gara, J.P. and Humphreys, H. (2008). Biofilm and the role of the ica operon and aap in Staphylococcus epidermidis isolates causing neurosurgical meningitis. Clinical Microbiology and Infection, 14(7): 719-722.
Suzuki, T., Kawamura, Y., Uno, T., Ohashi, Y. and Ezaki, T. (2005). Prevalence of Staphylococcus epidermidis strains with biofilm-forming ability in isolates from conjunctiva and facial skin. American Journal of Ophthalmology,140(5): 844-850
Vasudevan, P., Mohan-Nair, M.K., Annamalai, T. and Venkitanarayanan, K.S. (2003). Phenotypic and genotypic characterization of bovine mastitis isolates of Staphylococcus aureus for biofilm formation.Veterinary Microbiology, 92(1): 179-85.
Zmantar, T., Kouidhi, B., Miladi, H., Mahdouani, K. and Bakhrouf, A. (2010). A microtiter plate assay for Staphylococcus aureus biofilm quantification at various pH levels and hydrogen peroxide supplementation. New Microbiology, 33(2): 137-145.
