Enrofloxacin, Linco-Spectin ve Florfenicol antibiyotiklerinin BHK-21 hücre kültürü ve FMD 146S virus partikülü-infektif titreleri üzerine etkilerinin belirlenmesi

Yıl 2020, Cilt: 31 Sayı: 1, 7 - 19, 07.06.2020

Banu Bayri Özbilğe , Veli Gülyaz , Neslihan Taşçene , Şükran Yılmaz , Yasemin Gültekin , Ayşenur Ulusoy , Mustafa Hasöksüz , Gonca Öztap

https://doi.org/10.35864/evmd.729801

Cited By: 1

Öz

Abstract: Enrofloxacin, linko-spectin and florphenicol antibiotics were intended to be used in the BHK-21 An30 cells and the foot and mouth disease virus (FMDV) culture during the vaccine production process. Methylthiazolyldiphenyl-tetrazolium bromide (MTT) and cell cultures assays were performed in the BHK-21 An30 cells treated with the antibiotics that took place in this study to determine the concentration that inhibits cell proliferation and adverse effects on cell morphology. Virus cultures were performed by inoculating of FMDV serotypes (A/TUR/11, O/TUR/07, Asia-1/TUR/15) to the treated cell cultures with the antibiotics. 146S and infective tites of the obtained virus suspensions were determined. The non-toxic upper limit was determined to be 15μg /ml for enrofloxacin and 300μg/ml for linco-spectin in terms of cell morphology and cell numbers versus positive control (gentamycin, penicillin-streptomycin) and negative control (antibiotic-free medium) as a result of MTT and cell culture tests on BHK cells. It was observed that Florfenicol also inhibited cell proliferation and induced cell degeneration, even at a concentration of 5μg/ml. The mean 146S values of FMD virus cultures containing enrofloxacin
and linco-spectin were determined to be 0.49, 0.46, ; 0.53, 0.47 and 0.30, 0.28µg/ml for serotype A/TUR/11, O/TUR/07 and Asia-1/TUR/15 respectively. The mean values of the infective titres were 10^7,04, 10^7,25, 10^6,04, 10^6,59 and 10^7,26, 107,6 pfu/ml. for serotype A/TUR/11, O/TUR/07, and Asia-1/TUR/15 respectively. In the control group used gentamycin, penicillin-streptomycin and antibiotic-free medium, the mean 146S FMD virus particles were obtained as 0.51, 0.50, 0.50; 0.52, 0.55, 0.52 and 0.36, 0.33, 0.30 µg/ml for A/TUR/11, O/TUR/07 and Asia-1/TUR/15 respectively. The mean values of the FMD infective virus titres were 10^7,35, 10^7,40, 10^7,11; 10^6,24, 10^6,46, 10^6,62, and 10^7,70, 10^7,75, 10^7,77 pfu/ml for A/TUR/11, O/TUR/07 and Asia-1/TUR/15 respectively.
As a result, the infective FMDV titer and 146S results obtained in the control group (gentamicin, penicillin-streptomycin) and FMD virus cultures using Enrofloxacin (15µg / ml) and linco-spectin (300µg / ml) were very close to each other. According to these data, it was concluded that enrofloxacin and linco-spectin can be used up to the upper limit in the BHK-21 An30 cell and FMD virus cultures. However, florfenicol should not be used in cell and virus cultures.

Anahtar Kelimeler

Antibiyotik, BHK-21 An30, hücre kültürü, kontaminasyon, şap virüsü

Destekleyen Kurum

Tarımsal Araştırmalar ve Politikalar Genel Müdürlüğü

Proje Numarası

TAGEM/HSGYAD/15/A02/P02/58

Teşekkür

Bu projenin gerçekleştirilmesinde katkılarından dolayı Şap Enstitüsü Müdürü Dr. Nedret ÇELİK ve Dr. Can Çokçalışkan'a Teşekkür ederiz

Determination of the effects of Enrofloxacin, Linco-Spectin and Florphenicol antibiotics on BHK-21 cell culture and FMD 146S Virus particles-infective titers

Öz

Abstract: Enrofloxacin, linko-spectin and florphenicol antibiotics were intended to be used in the BHK-21 An30 cells and the foot and mouth disease virus (FMDV) culture during the vaccine production process. Methylthiazolyldiphenyl-tetrazolium bromide (MTT) and cell cultures assays were performed in the BHK-21 An30 cells treated with the antibiotics that took place in this study to determine the concentration that inhibits cell proliferation and adverse effects on cell morphology. Virus cultures were performed by inoculating of FMDV serotypes (A/TUR/11, O/TUR/07, Asia-1/TUR/15) to the treated cell cultures with the antibiotics. 146S and infective tites of the obtained virus suspensions were determined. The non-toxic upper limit was determined to be 15μg /ml for enrofloxacin and 300μg/ml for linco-spectin in terms of cell morphology and cell numbers versus positive control (gentamycin, penicillin-streptomycin) and negative control (antibiotic-free medium) as a result of MTT and cell culture tests on BHK cells. It was observed that Florfenicol also inhibited cell proliferation and induced cell degeneration, even at a concentration of 5μg/ml. The mean 146S values of FMD virus cultures containing enrofloxacin and linco-spectin were determined to be 0.49, 0.46, ; 0.53, 0.47 and 0.30, 0.28µg/ml for serotype A/TUR/11, O/TUR/07 and Asia-1/TUR/15 respectively. The mean values of the infective titres were 10^7,04, 10^7,25, 10^6,04, 10^6,59 and 1^07,26, 10^7,6 pfu/ml. for serotype A/TUR/11, O/TUR/07, and Asia-1/TUR/15 respectively. In the control group used gentamycin, penicillin-streptomycin and antibiotic-free medium, the mean 146S FMD virus particles were obtained as 0.51, 0.50, 0.50; 0.52, 0.55, 0.52 and 0.36, 0.33, 0.30 µg/ml for A/TUR/11, O/TUR/07 and Asia-1/TUR/15 respectively. The mean values of the FMD infective virus titres were 10^7,35, 10^7,40, 10^7,11; 10^6,24, 10^6,46, 10^6,62, and 10^7,70, 10^7,75, 10^7,77 pfu/ml for A/TUR/11, O/TUR/07 and Asia-1/TUR/15 respectively. As a result, the infective FMDV titer and 146S results obtained in the control group (gentamicin, penicillin-streptomycin) and FMD virus cultures using Enrofloxacin (15µg / ml) and linco-spectin (300µg / ml) were very close to each other. According to these data, it was concluded that enrofloxacin and linco-spectin can be used up to the upper limit in the BHK-21 An30 cell and FMD virus cultures. However, florfenicol should not be used in cell and virus cultures.

Anahtar Kelimeler

Antibiotic, BHK cell culture, contamination, FMD virus

Proje Numarası

TAGEM/HSGYAD/15/A02/P02/58

Kaynakça

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