Egypt. J. Plant Prot. Res. Inst. (2019), 2 (4): 617 - 723 Egyptian Journal of Plant Protection Research Institute www.ejppri.eg.net Potential antimicrobial activity of different types of Libyan honeys Abouzeid, A.S. 1; Reham, A.M. El-Wassef2 ; Sarah, H. El-Dereny3 ; Khaled, M.A. Abdel- Hameed3 and Mohamed, S. Younis³ ¹Entomology Deparment, Faculty of Science, Ain Shams University. ² Economic Entomology Department, Faculty of Agriculture, Cairo University. ³ Plant Protection Research Institute, Agricultural Research Center, Dokki, Giza, Egypt. ARTICLE INFO Abstract: Article History Honey exhibits antimicrobial activity against a wide range Received: 21/ 10 / 2019 of bacteria. The aim of the present work was to evaluate the Accepted: 18 / 11 /2019 antimicrobial effects of the Libyan honeys harmal (Peganum _______________ harmala L.) , red camphor (Cinnamomum camphora), white Keywords camphor (Eucalyptus globule), sarou (Cupressus sempervirens), Honey, antimicrobial athl (Tamarix aphylla ) and kharoub (Ceratonia silique ) on activity, bacteria , Staphylococcus aureus, Bacillus subtilis, Klebsiella pathogens and Libya. pneumoniae, Enterococcus faecalis, Pseudomonas aeuroginosa, Escherichia coli, Bacteriods spp., Sarcina spp. and Candida albicans. Pathogens exhibited different sensitivities towards the honey samples. The results showed that C. camphora inhibited seven out of the nine tested microorganisms followed by T. aphylla honey, which inhibited six of them. The lowest effects were shown by P. harmala and C. semperviren honeys, where they only inhibited four different types of the tested microorganisms. Introduction Honey is a complex natural food α-tocpherol (Ferreres et al.,1993). Honey produced from the honey bee Apis mellifera contains at least 181 components (White, L. (Hymenoptera: Apidae) feeding on plant 1975). nectar of blossoms, exudates of trees and The use of honey for the treatment of plants or from honey bees feeding on diseases and wounds has been mentioned honeydew produced by hymenoptran insects. since ancient time (2100-2000 BC), where Honey is a saturated solution of sugar of 31% Aristotle (384-322 BC) described pale honey glucose and 38% fructose and its colour and for sore eyes and wounds (Mandal and flavor vary considerably depending on it Mandal, 2011 and Vallianou et al. , 2014). botanical and geographical origin (Gheldof et Microorganisms such as Staphylococcus al., 2002) and of a moisture content of about aureus, Staphylococcus epidermis, 17.7% (Nagai et al., 2006). In addition to Micrococcus luteus, Streptococcus uberis, minor component of phenolic acids, Enterococcus faecalis, Pseudomonas flavonoids, glucose oxidase, catalase, aeruginosa, Escherichia coli and Klebsiella ascorbic acid, carotenoids, organic acids and pneumonia are frequently isolated from 717 Abouzeid et al. , 2019 human and animal skin wounds (Vuong and ulcers and the causative agent in many cases Otto, 2002; Nasser et al., 2003; Halcon and of dyspepsia has raised the possibility that the Milkus, 2004; Altoparlak et al., 2005 and therapeutic action of honey for symptoms of Basualdo et al., 2007). dyspepsia may be due to Manuka honey’s The healing effect of honey could be antibacterial properties. Somal et al. (1994) due to its physical and chemical properties demonstrated that after an incubation period (Rusell et al., 1990 and Snow and Manley- of 72 h.,5% Manuka honey completely Harris, 2004) and to its antioxidant and prevent the growth of H. pylori (the causative antimicrobial activity (Martos et al., 2000; organism of stomach ulcers). Similar invitro Escuredo et al., 2012; Vandamme et al., antimicrobial results of Manuka honey was 2013; Isidorov et al., 2015; Francine et al., reported against Campylobacter spp. (Lin et 2016; Almasaudi et al., 2017 and Leyva- al., 2009). Jimenez et al., 2019). Honey acts as an The aim of the present work was to effective thermal insulator and a protective evaluate the antimicrobial effects of the biofilm (Black and Costerton, 2010). Its Libyan honeys Harmal (Peganum harmala), antimicrobial activity is connected with its red camphor (Cinnamomum camphora), osmotic pressure which draws fluid from white camphor (Eucalyptus globule), sarou wounds, decreasing tissue edema (Molan, (Cupressus sempervirens), athl (Tamarix 2001). A possible reason for its activity aphylla) and Kharoub (Ceratoniasilique) on depends on its ability to generate hydrogen Staphylococcus aureus, Bacillus subtilis, peroxide by the bee derived enzyme glucose Klebsiella pneumoniae, Enterococcus dehydrogenase (Saleh et al., 2011). The faecalis, Pseudomonas aeuroginosa, strength of honey hydrogen peroxide is much Escherichia coli, Bacteriods spp., Sarcina lower than pharmacologic hydrogen spp. and Candida albicans. peroxide, causing no damaging to the healing Materials and Methods environment of a wound (Bang et al., 2003). The present investigation was carried Wound size is affected by pH value, Gethin out at the Beekeeping Research Section, et al. (2008) found that honey reduces the Plant Protection Research Institute, Giza, wound's pH and every 1 % reduction in pH is Egypt. associated with a 1 % reduction in wound 1.Honey samples: size. For wounds contaminated by Six types of Libyan honeys of mono methicillin-resistant Staphylococcus aureus, and multi-floral source were collected from Manuka honey supported better wound selected beekeepers during the harvesting healing than antibiotics (Gethin and periods and from local markets in Western Cowman, 2008). There are several studies Libya. The honeys of mono-floral source indicating the effectiveness of honey in were harmal (P. harmala, red camphor treating burns (Molan, 2001; Subrahmanyam, (C.camphora), white camphor (E. globule), 1991 and Subrahmanyam et al., 2001). sarou (C. sempervirens), athl (T. aphylla) and Sukur et al. (2011) studied the kharoub (C. silique). Honey samples were effectiveness of Tualang honey in healing kept in dark at room temperature prior to full-thickness burn wounds in rats. They analysis. The samples were investigated found that topical application of honey on microscopically to determine their containing burn wounds contaminated with of pollen grain types (Table, 1). Pseudomonas aeruginosa and Acinetobacter 2.Bacterial strains: baumannii give better results of healing Bacterial strains and C. albicans were compared with other treatments. kindly donated by the Microbial Genetic The finding that the bacterium, Department, Genetic Engineering and Helicobacter pylori is a cause of stomach 718 Egypt. J. Plant Prot. Res. Inst. (2019), 2 (4): 717 - 723 Biotechnology Division, National Research control are shown in Table (2). It was Center, Giza, Egypt. observed that all honey samples inhibited the 3. Assay of antimicrobial activity: growth of C. albicans with different degrees, Antimicrobial activity of honey where P<0.001. Bacteroids spp. was the most samples was determined by the disc diffusion resistant bacteria, where it was only inhibited method (Collins et al., 1995). A by C.camphora honey with an inhibition concentration of 20% of each kind of honey zone of only 5.0+0.67 mm. Except C. silique in distilled water was prepared in clean sterile honey all honey samples affect the growth of test tube and kept in refrigerator at 4oC to be E. faecalis with different degrees. B. subtilis used for microbiological test. was moderately inhibited by T. aphylla and 4.Preparation of the microbial culture: C. silique honeys with inhibition zones of The tested organisms were inoculated 11.33+0.57 and 11.66+0.57 mm, in the appropriate liquid media and incubated respectively. C. camphora inhibited seven at 37 oC for 24 h. The microbial culture was out of the nine tested microorganisms used for the preparation of seed layer by followed by T. aphylla honey, which inoculating the agar medium with 2% (v/v) of inhibited six of them.The lowest effects were the microbial culture, thoroughly mixed and shown by P.harmala and C. semperviren immediately used as the seed layer of plates. honeys, where they only inhibited four 5.Preparation of plates: different types of the tested microorganisms. The appropriate agar medium was Escherichia coli, P. aeruginosa and distributed at the rate of 7 ml portion in petri Bacteriods spp. were found to be resistant to dishes. After solidification 5 ml of the seeded the antibiotic tetramycine (+ve control), agar was distributed over the surface of the while 20% sucrose sugar solution (-ve base layer and left for 15 min to solidify. The control) had no inhibitory effect on all previously prepared filter paper discs (each bacterial strains. disc was moistened with exactly 0.05 ml of The antimicrobial activity of honey is the diluted honey) placed side down on the mainly contributed to the high osmolarity and seeded agar and gently pressed with a tip of acidity. In addition, hydrogen peroxide, sterile forceps. Discs were placed volatiles, organic acids, flavonoids, phenolic symmetrically around the center of the dish. compounds, wax, pollen, propolis are Plates were incubated at 37 oC for 24 hours. important factors that provide antimicrobial for P. aeruginosa and M. leutus, plates were properties to honey. Shin and Ustunol (2005) incubated at 30 oC. Antimicrobial activity stated that the sugar composition of honeys was determined measuring the diameter of from different floral source are responsible inhibition zones around the discs to the for the inhibition of various intestinal nearest mm (Table, 2). bacteria. According to Moumbe et al. (2013)