Probiyotik Etkili Bifidobacterium longum ATCC 15707’nin, Staphylococcus aureus ATCC 29213’e İn-vitro Etkisinin Real-Time PCR Yöntemi ile İncelenmesi

Staphylococcus aureus, kommensal bir bakteri olarak hem insanla birlikte yaşayan, hem de uygun fırsatı bulduğunda ciddi fırsatçı enfeksiyonlara neden olabilen önemli bir patojendir. Bifidobacterium cinsi bakteriler probiyotik olarak kullanılan ana mikroorganizmalardır. Bu cinslere ait çok sayıda türün, konağın sağlığını iyileştirmede güvenli ve etkili olduğu bildirilmiştir. Bizim çalışmamızın amacı, probiyotik etkili Bifidobacterium longum ATCC 15707’nin, Staphylococcus aureus ATCC 29213’e suşları üzerine etkisinin Real-Time PCR yöntemi ile araştırmaktır. 3 tane brain–heartin fusion broth (Oxoid) besiyeri içeren tüp hazırlandı. Tüm tüplere 103 CFU/mL olacak şekilde Staphylococcus aureus ATCC 29213 kökeninden ilave edildi. 1. tüp kontrol olarak kalırken, 2. tüpe 103 CFU/mL olacak şekilde, 3. tüpe de 106 CFU/mL olacak şekilde Bifidobacterium longum ATCC 15707 kökeni ilave edildi. İnkübasyonun 6., 12., 24. saatlerinde tüplerden alınan numunelerden DNA izolasyonu yapıldı ve bu numunelerden LightCycler 480 sisteminde Real-Time PCR ile kantitatif olarak S. aureus miktarları saptandı. Birinci tüpte 6. saatte 523.333+8993 kopya/mL düzeyinde saptanan S. aureus, farklı tüplerde sırasıyla 103 ve 106 CFU/mL miktarlarında B. longum kökeni ile birlikte inkübe edildiğinde bu miktarların düştüğü olduğu bulundu. Sonuç olarak, çalışmamızda, probiyotik etkili Bifidobacterium longum’un, Staphylococcus aureus’un in vitro ortamda üremesini etkilediği ve bakteri sayısının etki düzeyini arttırdığı saptanmıştır. Probiyotiklerin etkilerinin gnotobiyotik hayvan modellerinde veya daha kapsamlı çalışmalarla incelenerek, ilaç dirençlerinin çok yoğunlaştığı günümüzde, önemli patojenlere karşı antimikrobiyal olarak yarar sağlayabileceği düşüncesindeyiz.

Anahtar Kelimeler:

Bifidobacterium longum, Staphylococcus aureus, Real-Time PCR

Investigation of In vitro Effects of Probiotic Bifidobacterium longum ATCC 15707 on Staphylococcus aureus ATCC 29213 by Real-Time PCR

Staphylococcus aureus is an important pathogen which is both a commensal bacterium living with humans and can also cause serious infections when it has the opportunity. Bifidobacterium species are the main microorganisms used as probiotics. It has been reported that many species of this genus are safe an effective in improving the health of the host. The aim of this study was to investigate the effect of probiotic Bifidobacterium longum ATCC 15707 strain on Staphylococcus aureus ATCC 29213 strain by using with Real-Time PCR method. Three tubes containing brain-heartin fusion broth (Oxoid)were prepared. Staphylococcus aureus ATCC 29213 was inoculated into two tubes at a concentration of 103 CFU/mL while the one of the tubse remained as control. Bifidobacterium longum ATCC 15707 strain was added into the second and third tubes at a concentration of 103 CFU/mL and 106 CFU/mL respectively. DNA isolation was performed from the samples taken from tubes at 6th, 12th and 24th hours of incubation and quantitative S. aureus amounts were determined by Real-Time PCR by using LightCycler 480 system. S. aureus, amounts was found as 523.333+8993 copies/mL at the 6th hour in the first tube, while we found that the number of S. aureus was reduced in the tubes added with B. longum strains for 103 and 106 CFU/mL, respectively. In conclusion, B. longum, which is considered as a probiotic, affected the growth of Staphylococcus aureus in-vitro and the reducing effect of B. longum can increase along with the number. The effects of probiotics can be investigated in gnotobiotic animal models or in more comprehensive studies and we think that using B. longum can be beneficial against important pathogens such as Staphylococcus aureus in order to prevent antibiotic resistance.

Keywords:

Bifidobacterium longum, Staphylococcus aureus, Real-Time PCR,

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