Biofilm Production in Borehole Water Sources and their Susceptibility to Antibiotics and Antibiofilm Effect of Noni Foliar Extracts

Lawrence O Amadi, Faith T, Emeka, Grace I, Lekia
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Abstract

Background: Bacteria produce biofilm in a variety of systems; water, implant devices in humans and living tissues. They can be persistent and express virulent factors, increased resistance to antimicrobials and human immunity. Plant-based extracts have shown promising outcomes in inhibiting quorum sensing system and modulation of biofilm formation with lesser side effects. Thus, foliar extracts of Noni plant was the subject of such trial as an antibiofilm/antibacterial agent.  Objective: Investigate borehole water sources from three communities in Port Harcourt Local Government Area (PHALGA) for biofilm producers (BPs), antibiogram as well as antibiofilm effect of Noni (Morinda citrifolia) foliar extracts. Method: Microbiological protocols were adopted to isolate and identify bacteria whereas Congo red agar was used to detect biofilm producers (BPs). Antibiogram of BPs were done (with various antibiotics) as well as antibiofilm/antibacterial effect of Noni foliar extracts. Results: Bioassays detected the biofilm producers (BPs) as Staphylococcus aureus, Klebsiella pneumoniae, Bacillus and Serratia species. The bioburden were 55.0±2.5CFU/mL and 51.0±0.2CFU/mL for total heterotrophic bacteria and coliform counts from Ọbaziolu and Rumuelechi communities respectively.  Rumuelechi borehole water had the highest percentage relative abundance of S. aureus (80%) and  Bacillus (66.7%) whereas Ọbaziolu had Serratia (66.7%) and negative/zero for S. aureus and Ọrora had the least of all the isolated BPs. Antibiogram data revealed that all the BPs were 100% resistant to Cloxacillin and Cotrimoxazole but showed high rate of varying susceptibility to other antimicrobials; S. aureus depicted (100%) susceptibility to Gentamicin and Streptomycin; Bacillus to Tetracycline; Serratia to Augmentin and K. pneumoniae to Gentamicin. All the BPs showed multidrug resistance (MDR) with Serratia and K. pneumoniae being resistant to 6 and Bacillus and S. aureus 5-antibiotics respectively. Furthermore, susceptibility test on these BPs indicated that inhibitory activity of Noni foliar extracts were concentration-dependent. At a concentration of 3000mg/mL methanolic extract (MtE) showed the largest inhibition zone of 20.00mm against B. subtilis and aqueous extract (AqE) 15.00mm on Serratia species and such phenomenon is indicative of broad spectrum activity. Chloramphenicol (control) had the overall largest inhibition zones on the BPs except on B. subtilis. The susceptibility of these crude phytocompounds to BPs promises to be a novel and an alternative natural agent to synthetic antibiofilm products. Conclusion: Biofilm producers were detected in borehole water supply in three communities in Port Harcourt Local Government Area. The predominant BPs were Staphylococcus aureus, Bacillus and Serratia species. The BPs showed multi-antibiotic resistance with the Gram negative bacteria being resistant to more drug-types. Crude phytocompounds of Noni foliar extracts demonstrated broad spectrum activity with promising prospects as likely therapeutic option against biofilm-based infections. Additionally, the degree of multidrug resistance (MDR) depicted by these BPs to conventional antimicrobials is worrisome and demands regular monitoring, environmental sanitation and good hygienic measures to mitigate public health hazards.
钻孔水源生物膜的生成及其对抗生素的敏感性及诺丽叶提取物的抗生物膜作用
背景:细菌在各种系统中产生生物膜;水,人体和活体组织中的植入装置。它们可以是持续性的,表达毒力因子,增强对抗菌剂的耐药性和人体免疫力。植物提取物在抑制群体感应系统和调节生物膜形成方面显示出良好的效果,而且副作用较小。因此,诺丽植物叶面提取物是作为抗菌膜/抗菌剂进行试验的对象。目的:调查哈科特港当地政府区(PHALGA) 3个社区水井水源中生物膜生成物(bp)、诺丽(Morinda citrifolia)叶提取物的抗生素谱及抗膜作用。方法:采用微生物学方法分离鉴定细菌,刚果红琼脂检测生物膜生产者(bp)。用各种抗生素对bp进行抗生素谱测定,并观察诺丽叶提取物的抗菌作用。结果:生物测定检测到的生物膜生产者(bp)为金黄色葡萄球菌、肺炎克雷伯菌、芽孢杆菌和沙雷菌。Ọbaziolu群落异养菌总数为55.0±2.5CFU/mL,鲁木乐菌群落大肠菌群总数为51.0±0.2CFU/mL。Rumuelechi井水中金黄色葡萄球菌和芽孢杆菌的相对丰度最高(80%)和66.7%,Ọbaziolu井水中金黄色葡萄球菌的相对丰度最高(66.7%),金黄色葡萄球菌为阴性或零,Ọrora井水中分离的bp最少。抗生素谱数据显示,所有bp对氯西林和复方新诺明均100%耐药,但对其他抗菌素的不同敏感性较高;金黄色葡萄球菌(100%)对庆大霉素和链霉素敏感;对四环素的芽孢杆菌;沙雷氏菌配奥格门汀肺炎克雷伯菌配庆大霉素。所有bp均显示多重耐药(MDR),其中沙雷氏菌和肺炎克雷伯菌分别对6种抗生素耐药,芽孢杆菌和金黄色葡萄球菌分别对5种抗生素耐药。此外,对这些bp的药敏试验表明,诺丽叶提取物的抑制活性具有浓度依赖性。在浓度为3000mg/mL时,甲醇提取物(MtE)对枯草芽孢杆菌的最大抑制带为20.00mm,水溶液提取物(AqE)对沙雷氏菌的最大抑制带为15.00mm,表明其具有广谱活性。除枯草芽孢杆菌外,氯霉素(对照)对bp的总体抑制区最大。这些天然植物化合物对bp的敏感性有望成为合成抗生素膜产品的一种新的天然替代剂。结论:哈科特港3个社区的井水中均检测到生物膜生产者。主要的bp是金黄色葡萄球菌、芽孢杆菌和沙雷菌。bp表现出多种抗生素耐药性,革兰氏阴性菌对更多类型的药物具有耐药性。诺丽叶提取物的粗化合物具有广谱活性,有望作为生物膜感染的治疗选择。此外,这些bp描述的对常规抗菌素的多药耐药程度令人担忧,需要定期监测、环境卫生和良好的卫生措施,以减轻公共卫生危害。
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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