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

Yıl 2018, Cilt: 7 Sayı: 2, 229 - 232, 27.12.2018

Akın Yiğin , Mehmet Demirci

https://doi.org/10.31196/huvfd.508999

Öz

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

Öz

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 6^th,
12^th and 24^th 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.

Anahtar Kelimeler

Bifidobacterium longum, Staphylococcus aureus, Real-Time PCR

Kaynakça

• Bone RC, 1994: Gram-positive Organisms and Sepsis. Arch Intern Med, 154, 26-34.
• Brown AF, Leech JM, Rogers TR, Mc Loughlin RM, 2013: Staphylococcus aureus colonization: modulation of host ımmune response and ımpact on human vaccine design. Frontiers in Immunology, 4, 507.
• Hungin APS, Mulligan C, Pot B, P. Whorwell P, Agréus L, Fracasso P, Lionis C, Mendive J, Philippart J.M, Rubin G. Winchester C, Wit N, 2013: Systematic review: probiotics in the management of lower gastrointestinal symptoms in clinical practice–an evidence-based international guide. Alim Phar&Therap, 38, 864-886.
• Kim MJ, Ku S, Kim SY, Lee HH, Jin H, Kang S, Li R, Johnston TV, Park MS, Ji Ge, 2018: Safety evaluations of bifidobacterium bifidum bgn4 and bifidobacterium longum borı. Inter Jl of Mol Sci, 19, 1422.
• Lazarenko L, Babenko L, Sichel LS, Pidgorskyi V, Mokrozub V, Voronkova O, Spivak M, 2012: Antagonistic action of lactobacilli and bifidobacteria in relation to Staphylococcus aureus and their influence on the immune response in cases of intravaginal staphylococcosis in mice. Prob and Antimicr Prot, 4, 78-89.
• Lee DK, Kim MJ, Ham JW, An HM, Cha MK, Lee SW, Park CI, Shin SH, Lee KO, Kim KJ, Ha NJ, 2012: In-vitro evaluation of antibacterial activities and anti-inflammatory effects of Bifidobacterium spp. Address in gacnevulgaris. Arch Pharm Res, 35, 1065-71.
• Lenoir-Wijnkoop I, Gerlier L, Roy D, Reid G, 2016: The clinical and economic impact of probiotics consumption on respiratory tract infections: projections for Canada. Plos One, 11, 11.
• Lkhagvadorj E, Nagata S, Wada M, Bian L, Wang C, Chiba Y, Yamashiro Y, Shimizu T, Asahara T, Nomoto K, 2010: Anti-infectious activity of synbiotics in a novel mouse model of methicillin-resistant. Microbiol Immunol, 54, 265-275.
• Muñoz-Quezada S, Bermudez-Brito M, Chenoll E, Genovés S, Gomez-Llorente C, Plaza-Diaz J, Matencio E, Bernal MJ, Romero F, Ramón D, Gil A, 2013: Competitive inhibition of three novel bacteria isolated from faeces of breast milk-fed infants against selected enteropathogens. Br J Nutr, 109, 63-69.
• Phoem AN, Chanthachum S, Voravuthikunchai SP, 2015: Applications of micro encapsulated bifidobacterium longum with eleutherine americana in fresh milk tofu and pineapple juice. Nutrients, 7, 2469-2484.
• Reddy PN, Srirama K, Dirisala VR, 2017: An update on clinical burden, diagnostic tools, and the rapeutic options of Staphylococcus aureus. Infectious Diseases, 10, 1-15.
• Silva AKS, Silva TRN, Nicoli JR, Vasquez-Pinto LMC, Martins FS, 2018: In-vitro evaluation of antagonism, modulation of cytokines and extra cellular matrix proteins by bifidobacterium strains. Lett Appl Microbiol, 67, 497-505.
• Tong SYC, Davis JS, Eichenberger E, Holland TL, Fowler VG, 2015: Staphylococcus aureus ınfections: epidemiology, pathophysiology, clinical manifestations, and management. Clincal Micro Rev, 28, 603-661.
• Toscano M, De Grandi R, Stronati L, De Vecchi E, Drago L, 2017: Effect of Lactobacillus rhamnosus hn001 and Bifidobacterium longum bb536 on the healthy gut microbiota composition at phyla and species level: a preliminary study. World Journal of Gastroenterolog, 23, 2696-2704.
• Wang H, Yan Y, Wang J, Zhang H, Qi W, 2012: Production and characterization of antifungal compounds produced by Lactobacillus plantarum IMAU10014. PLoS ONE, 7, 29452.
• Wei Y, Yang F, Wu Q, Gao J, Liu W, Liu C, Guo X, Suwal S, Kou Y, Zhang B, Wang Y, Zheng K, Tang R, 2018: Protective effects of Bifidobacterial strains against toxigenic Clostridium difficile. Fron in Micr, 9, 888.