Variations in nutritional profile of honey produced by various species of genus Apis

Abstract The medicinal attributes of honey appears to overshadow its importance as a functional food. Consequently, several literatures are rife with ancient uses of honey as complementary and alternative medicine, with relevance to modern day health care, supported by evidence-based clinical data, with little attention given to honey’s nutritional functions. The moisture contents of honey extracted from University of Veterinary and Animal Sciences, Lahore honey bee farm was 12.19% while that of natural source was 9.03 ± 1.63%. Similarly, ash and protein contents of farmed honey recorded were 0.37% and 5.22%, respectively. Whereas ash and protein contents of natural honey were 1.70 ± 1.98% and 6.10 ± 0.79%. Likewise fat, dietary fiber and carbohydrates contents of farmed source documented were 0.14%, 1.99% and 62.26% respectively. Although fat, dietary fiber and carbohydrates contents of honey taken from natural resource were 0.54 ± 0.28%, 2.76 ± 1.07% and 55.32 ± 2.91% respectively. Glucose and fructose contents of honey taken out from honeybee farm were 27% and 34% but natural source were 22.50 ± 2.12% and 28.50 ± 3.54%. Glucose and fructose contents of honey taken out from honeybee farm were 27% and 34% but natural source were 22.50 ± 2.12% and 28.50 ± 3.54%. Similarly, sucrose and maltose contents of farmed honey were 2.5% and 12% while in natural honey were 1.35 ± 0.49% and 8.00 ± 1.41% respectively. The present study indicates that such as moisture, carbohydrates, sucrose and maltose contents were higher farmed honey as compared to the natural honey. In our recommendation natural honey is better than farmed honey.

giant bee), A. cerana (Indian bee), A. florae (little bee) and A. andreniformis although A. mellifera (European bee) is a pioneered species (Waghchoure-Camphor and Martin, 2008). Though the foremost 22 constituents of honey are nearly alike in every honey samples, the accurate chemical assemblage and physical characteristics of natural honeys dissimilar depending upon the plant kinds wherever the bees feed (James et al., 2009;Omafuvbe and Akanbi, 2009;Ebenezer and Olugbenga, 2010).
Facts show that few types of honey comprise kynurenic acid that is a tryptophan metabolite with neuroactive action that can impart to its antimicrobic qualities (Beretta et al., 2007). Analyses of the physical and chemical properties of honey are important for the official recognition procedure which establishes honey worth (Salim et al., 2011). Present study is therefore planned to find out variations in physicochemical characteristics and nutritional profile of honey produced by various Apis species.

Materials and Methods
A total of 5 honey samples of local honey bee species were collected from natural hives in the vicinity of Pattoki city while that of farmed species from Model Honeybee Farm, Department of Wildlife and Ecology, University of Veterinary and Animal Sciences, Ravi Campus ( Figure 1). Physical features of these honey samples viz. taste, smell, color and pH were recorded and the samples were stored in plastic glass bottles at room temperature for further analysis.

Proximate analysis
The proximate content, i.e. protein, fat, dietary fiber, carbohydrate, water and ash were determined depend on the official analysis methods as of Association of Official Analytical Chemists (AOAC). Every trial was performed in triplicate. Protein content was find out through Kjeldahl method depend upon the overall nitrogen content as of the AOAC Official Method 991.20, 2005. The fat content was find out by means of acid hydrolysis process. The dietary fibers comprised of the total, soluble and insoluble fibers of honey samples. 5 g of honey sample was used to measure and ash contents according to the AOAC Official Method (AOAC, 2005). Carbohydrate value was calculated using following Formula 1 (Charrondiere et al., 2004).
The energy values for the honey samples were also calculated following Formula 2 (Charrondiere et al., 2004).

Glucose oxidase activity with peroxide test
The activity of glucose oxidase in honey samples was monitored for peroxide accumulation using Merckoquant

Introduction
Every insect species has its own function into the environment and major groups are Coleoptera (beetles), Lepidoptera (butterflies and moths) and Diptera (flies). Furthermore, the Hymenopterans, specially the bees, are the mainly efficient pollinators of crops (bee keeping for pollination in regarding 70% of the earth's planted crops) and various other flowering plants. The honeybees have the powerful attraction to make honey and the scattering of seeds is also recognized for the purpose of pollination. The highly significant role of honeybee is that they pollinate the plants ( Morse and Calderon, 2000;Donkersley et al., 2017). There are three types of honeybee such as Apis florae, Apis dorsata and Apis cerana which play significant role in pollination (Partap, 2011). These are innate and known as the local kinds of Pakistan. Bees are differentiated from wasps by means of the existence of stemmed, frequently plumose hairs, and the posterior basitarsi, which are extensive than the successive tarsal fragments. The proboscis is in wide-ranging elongated than the majority sphecoid wasps. Michener (2007) noted 17,533 species of bees globally, characterized through 443 genera and seven families. Honey is a sugary, thick liquid prepared via honey bees. The smell, color and flavor of honey depend upon the different types of flowers. Chua and Adnan (2014) documented monosaccharide's, disaccharides and 17 amino acids from honey. The fitness promoting traits of honey are mostly because of the existence of numerous metabolites consisting of folic acid, thiamine, biotin, niacin, tocopherol, polyphenols, phytosterols besides enzymes and co-enzymes. The details lying on the antioxidant, anti-bacterial, anti-1 fungal, and hepato-protective characteristics of honey are well filed. In standard, honey is a precious addition for fit people (Denisow and Denisow-Pietrzyk, 2016;Muhammad et al., 2016). Honey cannot be consider a entire food by human being nutritional standards, on the other hand it put forwards strength such as a nutritional addition (Mendes et al., 1998).The usage of honey such as foodstuff and drug from human has been extended times before (Crane, 1983). Honey is made worldwide and its international production is more or less 1.20 million tons per annum (Bogdanov et al., 2008). Carbohydrates are amongst most important honey ingredients and are present in the variety of turanose, sucrose, maltulose, isomaltose, maltose, disaccharides, glucose, fructose, and monosaccharides. It also includes oligosaccharides comprising the anderose and panose and enzymes such as: amylase, oxidase peroxide, catalase and acid phosphorylase.
Numerous explorations have been attained in order to find out associations amongst the carbohydrate sketches and the nectar supplies with the usage of multivariate study (Cotte et al., 2004;Devillers et al., 2004). Moreover, honey includes amino acids, trace vitamin B, Vitamin B6, Vitamin C, niacin, folic acid, minerals, iron, zinc and antioxidants (Ball, 2007;Buba et al., 2013). Natural honey is produced by bees if different Apis species. The prevailing honey producing bees go to the genus Apis, underneath the family Apidae. Apis is symbolized via five species in Indian areas, of which four are local species for example: A. dorsata (rock bee or test strip (no. 10011) from Merck, Germany as illustrated by Kerkvliet (1996) and outcomes were stated in milligram of hydrogen peroxide accumulation in a liter of sample solution for an hour at 20 °C.

Sugar analysis by HPLC
The major sugar content of honey samples such as fructose, glucose, sucrose and maltose were analyzed using HPLC system coupled to a refractive index detector. Amide column with the dimension of 3.5 μm, 4.6 × 150 mm was applied for the separation. The column was kept at 25 °C all over the analysis. Honey samples (0.5 g) were dissolved in de ionized water and vortexes strongly before sieved used for injection. The injection volume was 20 μl. The average sugar solutions comprised of a combination of fructose, glucose, sucrose and maltose were prepared by diverse concentrations ranging from 5 to 20 g/kg meant for calibration curve construction.

Statistical analysis
The collected record was focused to statistical software SAS 9.1 and Analysis of Variance (ANOVA) were implemented to note inter specific variations in nutritional profile of honey from various honeybee species.

Results
The outcomes of the nutritional profile of honey samples obtained from UVAS Model Honeybee Farm are mentioned in Table 1. Moisture, ash, protein, fat, dietary fiber and carbohydrate values were 12.19%, 0.37%, 5.22%, 0.14%, 1.99% and 62.26%, respectively. Similarly, glucose, fructose, sucrose and maltose were recorded 27%, 34%, 2.5% and 12% respectively. Nutritional profile of honey samples extracted from different sources is mentioned in Table 1. The moisture contents of honey extracted from UVAS honeybee farm was 12.19% while that of natural source was 9.03 ± 1.63%. Similarly, ash and protein contents of farmed honey recorded were 0.37% and 5.22%, respectively. Whereas ash and protein contents of natural honey were 1.70 ± 1.98% and 6.10 ± 0.79%. Likewise fat, dietary fiber and carbohydrates contents of farmed source documented were 0.14%, 1.99% and 62.26%. Although fat,

Conclusion and Recommendations
Glucose and fructose contents of honey taken out from honeybee farm were 27% and 34% but natural source were 22.50 ± 2.12% and 28.50 ± 3.54%. Glucose and fructose contents of honey taken out from honeybee farm were 27% and 34% but natural source were 22.50 ± 2.12% and 28.50 ± 3.54%. Similarly, sucrose and maltose contents of farmed honey were 2.5% and 12% while in natural honey were 1.35 ± 0.49% and 8.00 ± 1.41% respectively. The present study indicates that such as moisture, carbohydrates, sucrose and maltose contents were higher farmed honey as compared to the natural honey. In our recommendation natural honey is better than farmed honey.