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Original Article ANTIBACTERIAL and ANTIOXIDANT ACTIVITY OF Bulgarian Journal of Veterinary Medicine, 2020 ONLINE FIRST ISSN 1311-1477; DOI: 10.15547/bjvm.2020-0133 Original article ANTIBACTERIAL AND ANTIOXIDANT ACTIVITY OF ROYAL JELLY COLLECTED FROM GEOGRAPHICAL REGIONS WITH DIFFERENT CLIMATES IN THE NORTH OF IRAN F. HAJI MOHAMMAD1, H. KOOHSARI2 & S. H. HOSSEINI GHABOOS1 1Department of Food Sciences, Azadshahr Branch, Islamic Azad University, Azadshahr, Iran; 2Department of Microbiology; Azadshahr Branch, Islamic Azad University, Azadshahr, Iran Summary Haji Mohammad, F., H. Koohsari & S. H. Hosseini Ghaboos, 2020. Antibacterial and anti- oxidant activity of royal jelly collected from geographical regions with different climates in the north of Iran. Bulg. J. Vet. Med. (online first). Royal jelly is one of the most important bee products. The biological activities of royal jelly can be influenced by various factors such as geographic origin, climatic conditions, vegetation. This study was carried out to evaluate the antibacterial and antioxidant activity of royal jelly samples collected from beehives from different geographical regions including mountain, coastal and plain regions in northern Iran. Antibacterial activity of royal jelly samples against ten bacteria was determined using agar well diffusion method. The MIC and MBC of royal jelly samples were determined by the broth microdilution method. Folin-Ciocâlteu reagent and reaction with DPPH were used to determine the total phenolic content and antioxidant potential of royal jelly samples, respectively. The MIC of samples ranged from 0.78 to 12.5% and MBC from 3.12 to 50%. Samples collected from mountain regions showed the highest antibacterial activity with MIC for Gram-positive bacteria from 0.78 to 1.56% and for Gram-negative bacteria: from 1.56 to 3.12%. The total phenolic content and DPPH radical scavenging activity in royal jelly samples of the mountain region was significantly higher than those from the two regions with other climates. The results of this study indicated that the climate of the geographic region of sampling location had an effect on the antibacterial and antioxidant activity of royal jelly which may be due to differences in plant vegetation and the origin of the flowers of bees. Key words: antibacterial activity, antioxidant activity, geographical region, royal jelly INTRODUCTION Today, due to population growth and the tance and unwanted side effects of chemi- limitations of modern medicine, the ne- cal agents. cessity of research into introducing new The honeybee is one of the most useful antimicrobial agents of natural origin is and most developed insects. Honeybee, on strongly needed to reduce antibiotic resis- the one hand, performs pollination of Antibacterial and antioxidant activity of royal jelly collected from geographical regions with … crops, gardens, and pasture plants increas- pounds. Royal jelly composition varies ing the crop yields and vegetation residues greatly according to the season, weather in rangelands, and on the other hand pro- conditions, bee nutrition, and climatic duces honey. Besides honey, other pro- conditions of the geographical region (Sa- ducts such as royal jelly, propolis, pollen, batini et al., 2009; Fratini et al., 2016). wax have many benefits to humans. Carbohydrates make up about 30% of Royal jelly is one of the most impor- the dry weight of the royal jelly. The most tant bee products, also called King Ang- abundant sugars include fructose, glucose, bin and King Jelly, which is attributed to and sucrose. Other oligosaccharides such numerous biological activities, including as maltose, trehalose, melibiose, ribose, antibacterial and antioxidant ones. Royal and other sugars are also found in small jelly is a result of the secretion of the hy- amounts (Fratini et al., 2016). popharyngeal glands of the bees, pro- Proteins are up to 50% of the dry duced at the age of 6 to 16 days. As age weight of the royal jelly. The most impor- increases and activity changes, the secre- tant royal jelly protein compositions in- tion of these glands changes as well. All clude the family of major royal jelly pro- larvae are fed this substance in the first teins (MRJPs), antimicrobial peptides three days and after 3 days, those who (AMPs) including royalisin, apismin, become worker bees are fed honey and jelleines, royalactin, apolipophorin III-like pollen, and those to be converted into a protein, and glucose oxidase (Fratini et queen are fed only royal jelly. Royal jelly al., 2016). plays an important role in the growth of Lipids make up 3 to 19% of royal infants, completing the special sexual jelly’s dry weight. Almost 90% is made traits and the long life of the queen. Di- up of fatty acids, and the rest is from neu- gestive and milky glands in the Honey- tral fats, steroids, hydrocarbons, and phe- bees by the production of royal jelly plays nols. The most important fatty acid in the an important role in the evolution of the structure of royal jelly is the 10 hydroxy-2 hive larvae, especially the queen. It seems decanoic acid (10-HDA) which is an un- that this kind of food causes the funda- saturated fatty acid involved in the anti- mental differences between the working bacterial activity of the product. This fatty bees and the queen, so that the life of the acid is the most important marker of royal queen is more than fifty-two times the jelly quality. Also, octanoic acid at length of life of the worker bees, and in amounts lower than those of 10-HDA has term of size the queen is 23 times larger nutritional roles (Fratini et al., 2016). than the workers (Fujita et al., 2013). Various studies have been reported the In terms of physical properties, royal differences and diversity of antibacterial jelly is a concentrated and milky sub- activities of royal jelly samples collected stance with a pungent smell and specific from different geographical regions (Gar- sour taste (pH =3.44.5), with a density of cia et al., 2010; 2013; Garcia-Amoedo & 1.1 g/cm3, slightly soluble in water (Fratini de Almeida-Muradian, 2007). et al., 2016; Sadredini et al., 2016). The antioxidant activity of royal jelly Royal jelly contains 6070% water, can be attributed to phenolic compounds 1123% carbohydrates, 918% protein, phenolic acid, cinnamic acid, coumarin, 48% lipids, and 0.83% vitamins, min- isocoumarin, naphthoquinone, xanthan, eral salts and low molecular weight com- anthraquinone, flavonoids, and lignin. 2 BJVM, ××, No × F. Haji Mohammad, H. Koohsari & S. H. Hosseini Ghaboos Numerous studies have pointed to the Iran including mountain, coastal, and diversity of phenolic compounds and anti- plain. Until the experiments samples were oxidant activity of royal jelly samples kept at 18 oC in the freezer of the Micro- collected from geographical regions with biology Laboratory at Islamic Azad Uni- different climates in different parts of the versity of Azadshahr branch. Samples world (Nabas et al., 2014; Ceksteryte et from beehives in the mountain region of al., 2016; Balkanska et al., 2017; Ozkok the Shahkuh-e Sofla in Gorgan township & Silici, 2017; Balkanska, 2018). The (mountain), coastal regions of the Faraha- difference in the amount of phenolic com- bad in Sari township (coastal), and plain pounds in the royal jelly depends on fac- regions of the Daland in Ramian township tors such as the weather, the herbal source (plain) were collected. The geographical used by the bee, the season, the genetic and vegetation characteristics of the re- factors, environmental factors such as the gions are presented in Table 1. type of soil (Perez-Perez et al., 2013; Sa- Serial dilution of royal jelly samples dredini et al., 2016). was carried out in sterilised distilled water The present study aimed to investigate and dilutions were prepared, resulting in the antibacterial and antioxidant effects of final concentrations of 100%, 50%, 25%, royal jelly collected from beehives from 12.5%, 6.3%, 3.12%, 1.56%, 0.78% and three geographical regions with different 0.39% v/v of each of the royal jelly sam- climates in the north of Iran. ples. Bacterial strains MATERIALS AND METHODS In this study, the antibacterial activity of Royal jelly samples royal jelly samples was evaluated against 4 bacterial native isolates and 6 bacterial Royal jelly samples were collected from reference strains. The reference strains beehives in three geographical regions used in this study comprised three Gram- with a different climate in the north of Table 1. Characteristics of royal jelly samples and collection regions Sample name and collection Climatic, geographical and vegetation characteristics region Shahkuh-e Sofla village is 65 km south-east of Gorgan in Golestan province. The village is 2540 meters above sea level with an average annual rainfall of 584 mm. Shahkuh-e It is a mountainous region with cold climate and snowy winters. The area’s vege- Sofla tation includes broad-leaved trees, shrubs and bush. Its plant species are yew, (mountain) aras, juniper, pyro, maze, beech, hornbeam, sloe, barberry, lily, blackbird, and herbaceous and bush species of oxtongue, alfalfa, sage, and artemisia. Farahabad is a coastal town 25 km north of Sari in Mazandaran Province. It is 26 Farahabad meters below sea level. The climate in this region is temperate with abundant (coastal) annual rainfall. The vegetation of the region includes agricultural lands paddy and gardens contains citrus fruits such as oranges, tangerine, and orange. Daland city of Ramian environs is located 60 km east of Gorgan in Golestan Daland province. The city is 65 meters
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