Correlation between relative bacterial activity and lactate dehydrogenase gene expression of co-cultures in vitro

Abstract

The present study aims at correlating the relative bacterial activity with the H+ concentration and the ldh expression of caries-associated bacteria in co-cultures. Well plates were prepared with BHI medium and cultures of Lactobacillus paracasei and Fusobacterium nucleatum. Bacterial growth at 37°C was measured using a microplate-photometer before and after adding sucrose to the samples. Samples of co-cultures (n = 12) and single-species cultures (n = 3) were taken and pH was assessed. Real-time quantitative PCRs were applied targeting the 16S-gene, the 16S-rRNA, the ldh-gene, and the ldh-mRNA. For L. paracasei with sucrose, an increase in relative bacterial activity (62.8% ± 23.5% [mean, SE]) was observed, while F. nucleatum showed a clear decrease in relative bacterial activity (- 35.0% ± 9.6%). Simultaneously, the H+ concentration increased (1.15E-05mol*l-1 ± 4.61E-07mol*l-1). Consequently, a significant positive correlation was found between L. paracasei's relative bacterial activity and H+ concentration (Spearman rank correlation, r = 0.638; p = 0.002), while F. nucleatum exhibited a negative correlation (r = - 0.741; p ≤ 0.001). Furthermore L. paracasei with sucrose showed a moderate, but significant positive correlation between relative bacterial activity and ldh-expression (r = 0.307; p = 0.024). The relative bacterial activity after sucrose pulse showed a significant correlation not only to the acid production (H+ concentration) but also to ldh expression of L. paracasei. However, further research is required to confirm these findings in a mature biofilm in vivo.