In vitro evaluation of microbial D- and L-lactate production as biomarkers of infection

Paula Morovic, M. Gonzalez Moreno, Andrej Trampuz, S. Karbysheva
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Abstract

Mammalian cells produce and metabolize almost exclusively L-lactate, bacterial species have the capacity to produce both D-lactate and L-lactate. The aim of this study was to evaluate the intrinsic production of D- and L-lactate in the most common pathogenic microorganisms causing septic arthritis (SA) and periprosthetic joint infection (PJI) as a potential biomarker for the diagnosis of infection. Following microorganisms were grown according to ATCC culture guides and tested for production of D- and L-lactate: Staphylococcus aureus (ATCC 43300), Staphylococcus epidermidis (ATCC 35984), Enterococcus faecalis (ATCC 19433), Streptococcus pyogenes (ATCC 19615), Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), Cutibacterium acnes (ATCC 11827), and Candida albicans (ATCC 90028). Pathogens were inoculated in 8 ml of appropriate liquid media and incubated as planktonic or biofilm form in either aerobic, anaerobic or CO2 atmosphere up to 312 h. D- and L-lactate measurements were performed at different time points: 0, 6, 9, 12 and 24 h, then once per day for slow-growing pathogens. Samples were serially diluted and plated for colony counting. Liquid culture media without microorganisms served as a negative control. Production of D-lactate was observed in all tested microorganisms, whereas no L-lactate was detected in E. coli, P. aeruginosa, and C. albicans. Maximal concentration of D-lactate was produced by S. aureus (10.99 mmol/L), followed by E. coli (1.22 mmol/L), and S. epidermidis (0.48 mmol/L). Maximal L-lactate concentration was observed in S. pyogenes (10.12 mmol/L), followed by S. aureus (9.71 mmol/L), E. faecalis (2.64 mmol/L), and S. epidermidis (2.50 mmol/L). S. epidermidis bacterial biofilm produced significantly higher amount of D- and L-lactate compared to planktonic form (p = 0.015 and p = 0.002, respectively). Our study has demonstrated that the most common pathogenic microorganisms causing SA and PJI have the capability to generate measurable amounts of D-lactate in both planktonic and biofilm form, highlighting the practical value of this biomarker as an indicator for bacterial and fungal infections. In contrast to D-lactate, the absence of L-lactate production in certain tested bacteria, as well as in fungi, suggests that L-lactate is not eligible as a biomarker for diagnosing microbial infections.
作为感染生物标志物的微生物 D-和 L-乳酸盐生成的体外评估
哺乳动物细胞几乎只产生和代谢 L-乳酸盐,而细菌则既能产生 D-乳酸盐,也能产生 L-乳酸盐。本研究的目的是评估导致脓毒性关节炎(SA)和假体周围关节感染(PJI)的最常见病原微生物中 D-乳酸和 L-乳酸的内在生成量,以此作为诊断感染的潜在生物标记物。根据 ATCC 培养指南培养了以下微生物,并对其产生的 D-和 L-乳酸进行了检测:金黄色葡萄球菌(ATCC 43300)、表皮葡萄球菌(ATCC 35984)、粪肠球菌(ATCC 19433)、化脓性链球菌(ATCC 19615)、大肠杆菌(ATCC 25922)、铜绿假单胞菌(ATCC 27853)、痤疮杆菌(ATCC 11827)和白色念珠菌(ATCC 90028)。将病原体接种到 8 毫升适当的液体培养基中,在有氧、厌氧或二氧化碳环境中以浮游生物或生物膜形式培养长达 312 小时:0、6、9、12 和 24 小时,对于生长缓慢的病原体则每天测量一次。对样品进行连续稀释,然后进行菌落计数。不含微生物的液体培养基作为阴性对照。在所有受测微生物中都观察到了 D-乳酸的产生,而在大肠杆菌、铜绿假单胞菌和白僵菌中没有检测到 L-乳酸。金黄色葡萄球菌产生的 D-乳酸浓度最高(10.99 毫摩尔/升),其次是大肠杆菌(1.22 毫摩尔/升)和表皮葡萄球菌(0.48 毫摩尔/升)。化脓性链球菌的 L-乳酸浓度最高(10.12 毫摩尔/升),其次是金黄色葡萄球菌(9.71 毫摩尔/升)、粪大肠杆菌(2.64 毫摩尔/升)和表皮葡萄球菌(2.50 毫摩尔/升)。与浮游生物相比,表皮葡萄球菌生物膜产生的 D-乳酸和 L-乳酸量明显更高(分别为 p = 0.015 和 p = 0.002)。我们的研究表明,导致 SA 和 PJI 的最常见病原微生物在浮游生物和生物膜形态下都能产生可测量的 D-乳酸,这突出了该生物标志物作为细菌和真菌感染指标的实用价值。与 D-乳酸相反,某些受测细菌和真菌不产生 L-乳酸,这表明 L-乳酸不能作为诊断微生物感染的生物标记物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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