Antibacterial activity of deer musk and Ziziphus spina-christi against carbapebem resis-tant gram negative bacteria isolated from patients with burns and wounds

Abstract

Bacteria were isolated from 250 specimens obtained from patients attending the Plastic Reconstructive and Burn Surgery Hospital in the Sulaymaniyah (Kurdistan region) and the burn and wound care units of the Azadi Teaching Hospital (Kirkuk), Iraq. Gram-negative bacteria were isolated from 100 (40%) of the samples which identified by BD phoenix, 66 isolates (66%) were carbapenem-resistant species, Rapidec® Carba NP test and sensitivity tests revealed 17 different genera and species of carbapenem-resistant bacteria. The BD Phoenix system was used to evaluate the susceptibility test of the isolates to 18 different antibiotics. The number of isolates that exhibited resistance to the carbapenem antibiotics, ertapenem, imipenem and meropenem, was 40 (61%), 35 (54%) and 28 (42%) respectively. Meanwhile, the number of isolates resistant to the penicillin antibiotics, ampicillin, amoxicillin-clavulanic acid and piperacillin-tazobactam, was 61 (92%), 54 (82%) and 24 (36%) respectively. The isolates were also evaluated for their resistance to the cephalosporins, cefazolin, cefuroxime, ceftazidime, ceftriaxone, cefepime, and ceftolozane-tazobactam; the respective number of resistant isolates were 60 (91%), 60 (91%), 39 (59%), 48 (73%), 42 (64%) and 27(41%). In contrast, a fraction of Ziziphus spina-christi leaves that was extracted using ethyl acetate inhibited all of the carbapenem-resistant isolates studied. The zone of inhibition (ZoI) Ø was between 19 and 24 mm. The ZoI Ø of black deer musk was 11–19 mm. The minimum inhibitory concentration (MICs) of ethyl acetate extract of Ziziphus spina-christi leaves for Acinetobacter baumannii (PDR), CR-Citrobacter farmeri, CR-Escherichia coli, CR-Proteus mirabilis, CR-Pseudomonas fluorescens, CR-Escherichia vulneris, CR-Kluyvera ascorbata, CR-Pantoea agglomerans, CR-Pseudomonas putida and CR-Serratia marcescens, was 6.25 mg/mL. Meanwhile, the MIC for Acinetobacter calcoaceticus–baumannii complex, Citrobacter freundii, Enterobacter cloacae, Klebsiella pneumoniae, Morganella morganii, Pseudomonas aeruginosa and Stenotrophomonas maltophilia was 12.5 mg/mL. High-performance liquid chromatography (HPLC) was used to analyse the components of the ethyl acetate-extract of Ziziphus spina-christi leaves. The analysis found gallic acid (760.3 ppm/mL), caffeine (84.0 ppm/mL) and quercetin (2.2 ppm/mL); the total phenolic content was 846.5 ppm/mL. The evidence obtained from this study indicates that leaves of this plant (named sidr in the Middle East) have the potential to be used as a natural antibiotic against carbapenem-resistant Gram-negative bacteria. Thus, the leaves of this tree present an important opportunity in the development of novel therapeutic agents. The study found the MIC of deer musk for CR-Citrobacter farmeri, CR-Citrobacter freundii, CR-Enterobacter cloacae, CR-Escherichia vulneris, CR-Klebsiella pneumoniae, CR-Kluyvera ascorbata, CR-Morganella morganii, CR-Pantoea agglomerans, CR-Proteus mirabilis, CR-Pseudomonas fluorescens, CR-Pseudomonas putida and CR-Serratia marcescens to be 50 mg/mL. In contrast, the MIC of deer musk was 100 mg/mL for Acinetobacter baumannii (PDR), CR-Acinetobacter calcoaceticus–baumannii complex, CR-Escherichia coli, CR-Pseudomonas aeruginosa and CR-Stenotrophomonas maltophilia. The results of the gas chromatography–mass spectrometry (GC–MS) indicate that the primary constituents of musk were1,4,4-tetramethyltetralin, 7-acetyl-6-ethyl-1, diethyl phthalate and tonalid; their contribution to the whole ranged from 12.2% to 19.6%. Black musk exhibits considerable antibacterial activity, able to inhibit seventeen different species of carbapenem-resistant Gram-negative bacteria. The non-natural form of black musk can be used to scent detergent.