First study of Micromonospora echinospora isolation from a rocky site of Eastern Algeria and first report of its potential use in cementitious materials biohealing

M. Kara Ali, Asma Ait Kaki, Wahiba Kara Ali, Amina Bramki, Amel Benchabbi, Noreddine Kacem Chaouche
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Abstract

In recent years, bio-healing based on microbial induced carbonate precipitation in cracks has been widely exploited to improve concrete properties and thus increase its durability. In this context, the present study aims to explore the possibility of using nine bacterial isolates as biohealing agent of cementitious materials; these bacteria (coded B1 to B8) were obtained from the three different rocky sites, i.e., 795 L1 (limestone rock site), 795 L2 (ordinary soil site), and 812 (clayey site); situated in Oulad Rahmoun (Constantine-Algeria). The primary identification showed that eight isolates belonged to a Bacillus genus, and one isolate B8 developed particularly morphological characterized by branched and septate hyphae. The isolates B4 and B8 were selected for their ability to produce calcium carbonate in precipitate calcium (PC) liquid and solid media after five days of incubation. The addition of urea in PC liquid media accelerated the formation of calcite which appeared after 3 days of incubation. The growth curve of both selected isolates on LB liquid medium showed that B8 was characterized by a long exponential growth phase (up to 72 hours) compared to B4 which had an exponential phase up to 48 hours, which could explain the largest amount of calcite precipitate observed in the case of B8 cultivated in PC liquid medium. Results of the bio-healing experiment carried on cementitious batches showed that a beginning of a slight repair of the cracks from the 11th day was only observed in the case of the bacterium B8 but the bio-healing capacity was absent in the control. This bio-healing agent was identified as Micromonospora echinospora (NCBI-NO139612), according to N-Blast of 16S-DNA sequence. In conclusion, the present study is the first to isolate M. echinospora species from rocky sites; and to describe it as an efficient bacterial agent in concrete bio-healing applications.
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阿尔及利亚东部岩石遗址分离棘孢小单孢子虫的首次研究及其在胶凝材料生物愈合中的潜在应用的首次报道
近年来,基于裂缝中微生物诱导碳酸盐沉淀的生物修复技术被广泛应用于改善混凝土性能从而提高其耐久性。在此背景下,本研究旨在探索使用9种细菌分离物作为胶凝材料生物愈合剂的可能性;这些细菌(编码B1 ~ B8)分别来自3个不同的岩石样地,即795 L1(石灰岩样地)、795 L2(普通土壤样地)和812(粘土样地);位于Oulad Rahmoun(君士坦丁-阿尔及利亚)。初步鉴定表明,8株分离菌株属于芽孢杆菌属,其中1株B8菌丝呈分枝和分离形态。菌株B4和B8在沉淀钙(PC)液体和固体培养基中培养5天后均能产生碳酸钙。在PC液体培养基中添加尿素加速了方解石的形成,培养3天后方解石出现。两株菌株在LB液体培养基上的生长曲线表明,B8的指数生长期较长(可达72 h),而B4的指数生长期为48 h,这可以解释B8在PC液体培养基上培养时方解石析出量最大的原因。对胶凝批进行的生物愈合实验结果表明,从第11天开始,只有细菌B8开始轻微修复裂缝,而对照组没有生物愈合能力。该生物愈合剂经16S-DNA序列N-Blast鉴定为棘皮小单孢菌(Micromonospora echinospora, NCBI-NO139612)。综上所述,本研究是首次从岩石遗址中分离出棘孢支原体;并将其描述为混凝土生物愈合应用中的高效细菌剂。
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