Molecular Characterization of ESBL Gene in Citrobacter Spp and Antibacterial Activity of Omega-3 Against Resistant Isolates

Mayyada Farhan Darwees
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Abstract

Abstract This study aimed to investigated the prevalence and resistance pattern of different Citrobacter species phenotypically and genotypically to β-lactam and some most common antibiotics then evaluate the antibacterial activity of omega-3 extracted from flaxseed against isolates that harboring resistance genes. 19 Citrobacter isolates were isolated from100 stool and urine samples taken from patients attended to AL-Sadar Hospital during June-December 2016. Clinical samples were cultured on specific media, thereafter isolates were identified depending on morphological, biochemical characteristics and VITK-2. The results showed that the Citrobacter comprise 24% of isolated bacteria which divided into 11 (14.1%) were C. freundii, 5 (6.41%) C. kosaeri and C. farmeri were 3 (3.8%). The antagonistic activity was evaluated by observing a clear zone of inhibition growth, the results showed that all Citrobacter (100%) isolates were resistant to Ampicillin, cefoxitin and sensitive to Imipinim, also the isolates showed different degrees of resistant to β-lactam antibiotics initially. By confirmatory test the results observed 17/19 (89.4%) isolated were ESBL producers finally using PCR technique to detect bla–genes (blaCTX-M, OXA, SHV and Z). The results revealed that 14/17 (82.3) of potential ESBL producing Citrobacter were harbor one or more of ESBL genes they included 10 of C. freundii and 4 of C. koseri. The extraction of essential fatty acid semicarbazide (omega-3) from Linum usitatissium (Flax seed) were tested to evaluate their activity against resistant isolates, results explained broad spectrum antibacterial property of EFASC compounds against resistant bacteria. In conclusion, this study found increase prevalence of MDR Citrobacter spp as causative agents in clinical cases. Considering antibacterial activity of EFASC, it was observed highly activity against resistant pathogens deservedly, therefore attention must be paid to development their used as alternative antibiotics.
柠檬酸杆菌ESBL基因的分子特征及Omega-3对耐药菌株的抑菌活性
摘要本研究旨在研究不同柠檬酸杆菌种类对β-内酰胺和一些最常见抗生素的表型和基因表型的流行情况和耐药模式,并评价从亚麻籽中提取的omega-3对含有耐药基因的分离株的抑菌活性。从2016年6月至12月在AL-Sadar医院就诊的患者的100份粪便和尿液样本中分离出19株柠檬酸杆菌。临床样品在特定培养基上培养,然后根据形态、生化特征和VITK-2鉴定分离株。结果表明,分离菌中柠檬酸杆菌占24%,其中弗氏梭菌11种(14.1%),科氏梭菌5种(6.41%),法氏梭菌3种(3.8%)。结果表明,所有Citrobacter菌株(100%)均对氨苄西林、头孢西丁耐药,对亚胺平敏感,对β-内酰胺类抗生素均表现出不同程度的耐药。结果显示,17/19株(89.4%)菌株为ESBL产生菌,最终采用PCR技术检测bla基因(blaCTX-M、OXA、SHV和Z),其中14/17株(82.3)可能产生ESBL的Citrobacter含有一个或多个ESBL基因,其中C. freundii 10株,C. koseri 4株。从亚麻籽中提取必需脂肪酸氨基脲(omega-3),测定其对耐药菌株的抑菌活性,结果表明EFASC化合物对耐药菌具有广谱抗菌作用。总之,本研究发现临床病例中耐多药柠檬酸杆菌作为病原体的患病率增加。考虑到EFASC的抗菌活性,其对耐药病原菌具有较高的抗菌活性,因此应重视其作为替代抗生素的开发。
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