Investigation of Tetracycline Resistance of Lactic Acid Bacteria by in silico Genotypically in Whole Genome Sequences and Phenotypically

Öz

Lactic acid bacteria are used in industry as starter cultures or probiotics. In the report published by the European Food Safety Authority in 2021, it was emphasized that risk assessment should be carried out on the whole genome sequences of bacteria to be used in foods. Therefore, resistance gene research in lactic acid bacteria has gained importance. Because antibiotic resistance genes are likely to be transmitted to pathogenic bacteria in the intestinal tract or are likely to be ingested through foods containing lactic acid bacteria. Therefore, in this study, four lactic acid bacteria (Lentilactobacillus buchneri Egmn17, Levilactobacillus brevis Atlas17, Levilactobacillus namurensis Ozge01, Lactiplantibacillus plantarum Gmze16) isolated from different fermented foods and Lactiplantibacillus plantarum, a probiotic bacterium, were used. In the study, tetracycline, which has the most common antibiotic resistance among lactic acid bacteria, was selected. It was determined that 3 bacteria were moderately resistant (zone diameter 15-18 mm) (299v, Gmze16 and Egmn17) to tetracycline antibiotic and 2 bacteria were susceptible (zone diameter >19 mm) (Atlas17 and Ozge01). As a result of examining the whole genome sequences of lactic acid bacteria, it was seen that intermediate resistant bacteria have tetracyclinedependent antimicrobial resistance (AMR) genes tetA (MFS efflux pump) and tetO (ribosomal protection protein). In Levilactobacillus brevis Atlas17, while TetA protein was present, M→T change was observed in the 322nd amino acid sequence. In addition, the fact that the probiotic bacteria Lactiplantibacillus plantarum 299v has resistance genes also increases the risk of transferring these genes to pathogens in the intestine. Species that are susceptible to the phenotype, such as Levilactobacillus brevis Atlas17, which has been observed to have the tetA gene, are also likely to be able to pass it on to other bacteria when they have silent resistance genes. Therefore, it is important to examine genotype and phenotype together.

Anahtar Kelimeler

• • Tetracycline resistance
• • Lactic acid bacteria
• • Whole genome sequence
• • MFS efflux pump
• • tetO gene

Laktik Asit Bakterilerinde Tetrasiklin Direncinin Fenotipik ve Tüm Genom Dizilerinde in silico Genotipik Olarak Araştırılması

Öz

Laktik asit bakterileri, endüstride starter kültür veya probiyotik olarak kullanılmaktadırlar. European Food Safety Authority (EFSA) tarafından 2021 yılında yayımlanan raporda gıdalarda kullanılacak bakterilerin tüm genom dizileri üzerinden risk değerlendirmesi yapılması gerekliliği vurgulanmıştır. Bu nedenle, laktik asit bakterilerinde dirençlilik geni araştırmaları önem kazanmıştır. Çünkü antibiyotik direnç genlerinin bağırsak sisteminde bulunan patojen bakterilere aktarılma olasılığı vardır ya da laktik asit bakterilerini barındıran gıdalar aracılığıyla alınmaları olasıdır. Bu nedenle, çalışmada, farklı fermente gıdalardan izole edilen dört laktik asit bakterisi (Lentilactobacillus buchneri Egmn17, Levilactobacillus brevis Atlas17, Levilactobacillus namurensis Ozge01, Lactiplantibacillus plantarum Gmze16) ve probiyotik bir bakteri olan Lactiplantibacillus plantarum 299v suşu kullanılmıştır. Çalışmada, laktik asit bakterileri arasında en yaygın antibiyotik dirençliliği gözlenen tetrasiklin seçilmiştir. 3 bakterinin tetrasiklin antibiyotiğine orta derecede dirençli (zon çapı 15-18 mm) (299v, Gmze16 ve Egmn17) ve 2 bakterinin duyarlı (zon çapı >19 mm) (Atlas17 ve Ozge01) olduğu belirlenmiştir. Laktik asit bakterilerinin tüm genom sekanslarının incelenmesi sonucu, orta dirençli bakterilerin tetrasikline bağlı antimikrobiyal direnç (AMR) genlerinden tetA (MFS dışa atım pompası) ve tetO’ya (ribozomal koruma proteini) sahip oldukları görülmüştür. Levilactobacillus brevis Atlas17’de ise TetA proteini mevcutken 322. aminoasit sekansında M → T değişimi gözlenmiştir. Ayrıca probiyotik bakteri olan Lactiplantibacillus plantarum 299v’nin direnç genlerine sahip olması bu genlerin bağırsaktaki patojenlere aktarılma riskini de arttırmaktadır. tetA genine sahip olduğu gözlenen Levilactobacillus brevis Atlas17 gibi fenotipi duyarlı olan türler de sessiz dirençlilik genlerine sahip olduklarında bunu diğer bakterilere aktarabilmeleri olasıdır. Bu nedenle genotip ve fenotip birlikte incelenmesi önemlidir

Anahtar Kelimeler

• • Tetrasiklin direnci
• • Laktik asit bakterisi
• • Tüm genom sekansı
• • MFS dışa atım pompası
• • tetO geni

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