Mellifera Syriaca Research Articles

Effect of Apis mellifera syriaca Bee Venom on Glioblastoma Cancer: In Vitro and In Vivo Studies.

Glioblastoma multiforme (GBM) is a highly aggressive and fatal primary brain tumor. The resistance of GBM to conventional treatments is attributed to factors such as the blood-brain barrier, tumor heterogeneity, and treatment-resistant stem cells. Current therapeutic efforts show limited survival benefits, emphasizing the urgent need for novel treatments. In this context, natural anti-cancer extracts and especially animal venoms have garnered attention for their potential therapeutic benefits. Bee venom in general and that of the Middle Eastern bee, Apis mellifera syriaca in particular, has been shown to have cytotoxic effects on various cancer cell types, but not glioblastoma. Therefore, this study aimed to explore the potential of A. mellifera syriaca venom as a selective anti-cancer agent for glioblastoma through in vitro and in vivo studies. Our results revealed a strong cytotoxic effect of A. mellifera syriaca venom on U87 glioblastoma cells, with an IC50 of 14.32 µg/mL using the MTT test and an IC50 of 7.49 µg/mL using the LDH test. Cells treated with the bee venom became permeable to propidium iodide without showing any signs of early apoptosis, suggesting compromised membrane integrity but not early apoptosis. In these cells, poly (ADP-ribose) polymerase (PARP) underwent proteolytic cleavage similar to that seen in necrosis. Subsequent in vivo investigations demonstrated a significant reduction in the number of U87 cells in mice following bee venom injection, accompanied by a significant increase in cells expressing caspase-3, suggesting the occurrence of cellular apoptosis. These findings highlight the potential of A. mellifera syriaca venom as a therapeutically useful tool in the search for new drug candidates against glioblastoma and give insights into the molecular mechanism through which the venom acts on cancer cells.

Apis mellifera syriaca Venom Modulates Splenic Cytokines Levels in BALB/c Mice.

Bee venoms are well-known for their important biological activities. More specifically, the venom of Apis mellifera syriaca was shown to exhibit various biological effects, including antimicrobial effects. It is suggested that the anti-microbial effect of venom could be accompanied by an immunomodulatory response in the host favoring anti-inflammatory responses. Thus, in this work, we investigated, for the first time, the immunomodulatory effects of A. mellifera syriaca venom in mice. Firstly, it was found that this venom exhibited mild toxicity in BALB/c mice after intraperitoneal injection with an LD50 of 3.8 mg/kg. We then investigated its immunomodulatory effects by evaluating the splenic levels of both pro- and anti-inflammatory cytokines in mice by ELISA. Interestingly, at 1 mg/kg, A. mellifera syriaca venom induced a decrease in IFN-γ, TNF-α, IL-4, and IL-10 at 24h postinjection. At a higher dose (3 mg/kg), an increase in IFN-γ and IL-4 levels was observed, while the levels of TNF-α and IL-10 remained low compared to the control. Altogether, these preliminary data suggest that A. mellifera syriaca venom exhibits anti-inflammatory effects at a sublethal dose (1 mg/kg), while at a higher dose (3 mg/kg), it induces inflammatory effects.

Apis mellifera syriaca Venom: Evaluation of Its Anticoagulant Effect, Proteolytic Activity, and Cytotoxicity along with Its Two Main Compounds-MEL and PLA2-On HeLa Cancer Cells.

Bee venom (BV) is one of the most remarkable natural products that has been a subject of studies since ancient times. Recent studies have shown that Apis mellifera syriaca venom possesses antibacterial as well as cytotoxic effects on cancer cell lines. The venom contains a variety of bioactive molecules—mainly melittin (MEL) and phospholipase A2 (PLA2), as well as other compounds that are not well characterized. In this work, we continue the biological characterization of A. mellifera syriaca venom by testing its anticoagulant effect on human plasma using the prothrombin time (PT) test, as well as assessing its proteolytic activity. In addition, the cytotoxicity of the crude venom—and of its two main components, MEL and PLA2—was tested on HeLa cancer cell lines for the first time. The results obtained showed the capacity of A. mellifera syriaca venom to increase clotting time, thereby proving its anticoagulant effect. Moreover, the venom did not demonstrate a significant proteolytic activity unless administrated at concentrations ≥ 5 mg/mL. Finally, we showed that crude A. mellifera syriaca venom, along with MEL, exhibit a strong in vitro cytotoxic effect on HeLa cancer cell lines, even at low concentrations. In summary, our findings could serve as a basis for the development of new natural-based drug candidates in the therapeutic field.

EFFECTS OF HONEY BEE RACE AND SEASON ON PROPOLIS COMPOSITION

Honey bees collect the main material of propolis from the buds, leaves, branches, and barks or other botanical sources and mixed that resinous material with beeswax produced from abdominal exocrine glands, mandibular gland secretions, and pollen to produce propolis. The composition of propolis changes depends on multiple factors such as honey bee races, geographical locations, phytogeography, harvesting seasons, extraction methods and solvents. In this study, two different studies were conducted in two different locations in order to reveal the effect of race and season variables on the composition and antioxidant value of propolis. The effect of race factor was studied on three different honey bee races (Apis mellifera caucasica, Apis mellifera syriaca and Apis mellifera carnica) and two different ecotypes (Muğla ecotype and Yığılca ecotype) to investigate the effects of race factor on propolis composition in Central Anatolia by under the controlled conditions. The effect of seasonal change was determined by Yığılca ecotype of A. melifera anatoliaca in the apiary located in Yığılca, Düzce location. Studied samples were harvested by propolis trap between May and October. Total phenolic content, total antioxidant capacity and chemical profiles of propolis samples were determined using HPLC-UV. The obtained results showed that honey bee race and season have an effect on the antioxidant capacity and chemical composition of propolis.

Anadolu’daki Bal Arısı Biyoçeşitliliği Yok Olma Sürecinde mi?

This study was conducted through the geometric morphometric method by making a sampling as to represent seven geographical regions of Turkey and also all honeybee races that are reported to be found in Turkey. Nineteen landmarks on the wings of bees detected by using right front wings of worker bees were measured by the Bs200Pro program. According to the results of discriminant function analysis based on individual data, the true classification rate was found to be 54.1%. While the Marmara, Aegean, Eastern Anatolia and Black Sea Regions formed a strictly single group, the Southeastern Anatolia Region distinctly separated from other regions. The honey bee population of eastern and western parts of the Mediterranean Sea were divided into two, while the western part formed close groups with other regions, the eastern part got closer to the Southeastern Anatolia Region. The group center of the Central Anatolia Region partly separated from the other regions and formed a group at the edge. Beside the split of the Mediterranean Region as East and West, the samples taken from Southeastern Anatolian Region create a separate group supported the existence of Apis mellifera syriaca and Apis mellifera meda races in the South. The separation of the Central Anatolia Region supports the existence of Apis mellifera anatoliaca in Central Anatolia of Turkey. However, the examples taken from other regions nested together to create a group show that the honey bee biodiversity in Turkey has been affected by the migratory beekeeping and commercial queenbee activities in recent years.

The Relationships Between Propolis Collecting Capability and Morphometric Features of Some Honey Bee Races and Ecotypes in Anatolia

Propolis collecting capacity of the honey bee race, Apis mellifera L., distributed across Anatolia and Thrace regions of Turkey was investigated and correlated with morphometric characteristics. Thus, the propolis collecting behaviour of honey bee races and ecotypes naturally have been in Turkey, Apis mellifera caucasica, Apis mellifera carnica, Apis mellifera syriaca and Yigilca and Mugla ecotype of Apis mellifera anatoliaca were monitored. The mean yield of annual propolis was recorded as the following; Yigilca ecotype (111.6±27.5 g colony) A. m. caucasica (104±20.7 g colony), Mugla ecotype (103±34 g colony), A. m. carnica (91.16±17.6 g colony), and A. m. syriaca (74±6.4 g colony) in descending order. The highest propolis collecting activity was recorded for the Yigilca ecotype of A. m. anatoliaca and A. m. caucasica. Morphological features of honey bee samples were evaluated by classic morphometric technique to correlate propolis collecting capability and morphological features. Morphometric results of the present study showed that the largest wing and leg lengths belonged to Yigilca ecotype of A. m. anatoliaca and A. m. caucasica. Furthermore, Pearson correlation showed a significant relationship between some morphometric characteristics including the proboscis and mandibular sections, wing length (WL), wing width (WW), femur length (FL), tibia length (TL), basitarsus length (BL), basitarsus width (BW), and propolis collecting capability (P<0.05). Therefore, it seems that the enlargement of certain morphological properties with genetic tendency of the honey bee races and ecotypes, primarily the legs and wings, can lead to better propolis collecting capability.

Evaluation of antioxidant activity of bee products of different bee races in Turkey

Bee products with high bioactive properties have a very important place in human nutrition. Biochemical properties of these products such as honey, pollen, and propolis vary according to flora, season, bee race, and production methods. In this study, the biological activities of honey, bee pollen, and propolis samples from four bee races in Turkey (Apis mellifera caucasica, Apis mellifera anatoliaca, Apis mellifera syriaca, and Apis mellifera carnica) were examined. Besides the determining of total polyphenol and total flavonoid contents, the antioxidant capacity was investigated by three different assays: DPPH (1,1-diphenyl-2-picrylhydrazyl) radical-scavenging activities, CUPRAC (cupric reducing antioxidant capacity) and FRAP (ferric reducing antioxidant power). The rank order of antioxidant potencies was as follows in the four bee races: propolis > pollen > honey. The polyphenol content of the products was evaluated and Apis mellifera syriaca for honey, Apis mellifera anatoliaca for pollen, and Apis mellifera carnica for propolis were found higher than the others. When bee races were compared, it was seen that flora is a more effective factor than bee race for antioxidant capacity.

First Characterization of The Venom from Apis mellifera syriaca, A Honeybee from The Middle East Region.

Bee venom is a mixture of several components with proven therapeutic benefits, among which are anti-inflammatory, analgesic, and various cardiovascular conditions. In this work, we analyzed for the first time the proteomic content and biological properties of the crude venom from Apis mellifera syriaca, a honeybee from the Middle East region. Using high-performance liquid chromatography-tandem mass spectrometry, we evidence the venom contains phospholipase A2, hyaluronidase, mast cell-degranulating peptide, adolapin, apamin, and melittin. The latter was purified by solid phase extraction method (SPE) and tested in parallel with crude venom for biological activities. Precisely, crude venom—but not melittin—exhibited antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa strains. Alongside, hemolytic activity was observed in human blood subjected to the venom at high doses. A. mellifera syriaca venom displayed antioxidant activities, and not surprisingly, PLA2 catalytic activity. Eventually, the venom proved to exert antiproliferative effects against MCF-7 and 3T3 cancer cells lines. This first report of a new bee venom opens new avenues for therapeutic uses of bee venoms.

Appetitive reversal learning differences of two honey bee subspecies with different foraging behaviors.

We aimed to examine mechanistically the observed foraging differences across two honey bee, Apis mellifera, subspecies using the proboscis extension response assay. Specifically, we compared differences in appetitive reversal learning ability between honey bee subspecies: Apis mellifera caucasica (Pollman), and Apis mellifera syriaca (Skorikov) in a “common garden” apiary. It was hypothesized that specific learning differences could explain previously observed foraging behavior differences of these subspecies: A.m. caucasica switches between different flower color morphs in response to reward variability, and A.m. syriaca does not switch. We suggest that flower constancy allows reduced exposure by minimizing search and handling time, whereas plasticity is important when maximizing harvest in preparation for long winter is at a premium. In the initial or Acquisition phase of the test we examined specifically discrimination learning, where bees were trained to respond to a paired conditioned stimulus with an unconditioned stimulus and not to respond to a second conditioned stimulus that is not followed by an unconditioned stimulus. We found no significant differences among the subspecies in the Acquisition phase in appetitive learning. During the second, Reversal phase of the experiment, where flexibility in association was tested, the paired and unpaired conditioned stimuli were reversed. During the Reversal phase A.m. syriaca showed a reduced ability to learn the reverse association in the appetitive learning task. This observation is consistent with the hypothesis that A.m. syriaca foragers cannot change the foraging choice because of lack of flexibility in appetitive associations under changing contingencies. Interestingly, both subspecies continued responding to the previously rewarded conditioned stimulus in the reversal phase. We discuss potential ecological correlates and molecular underpinnings of these differences in learning across the two subspecies. In addition, in a supplemental experiment we demonstrated that these differences in appetitive reversal learning do not occur in other learning contexts.

Next-generation sequence data demonstrate several pathogenic bee viruses in Middle East and African honey bee subspecies (Apis mellifera syriaca, Apis mellifera intermissa) as well as their cohabiting pathogenic mites (Varroa destructor).

RNA viruses are associated with honey bee (Apis mellifera) colony losses in many parts of the world. Their consequences may be exacerbated when the ectoparasite mite Varroa destructor is present in hives. While evidence of pathogenic, viral-induced disease is abundant in western honey bees (Apis mellifera mellifera) from many parts of the world, less information exists regarding the pathogen load of Apis mellifera syriaca and Apis mellifera intermissa, honey bees from the Middle East and North Africa (MENA) that play substantial roles in regional beekeeping. Here, we used next-generation sequencing to evaluate the viral populations of these subspecies and their associated mites. We found that both A. m. syriaca and A. m. intermissa, as well as the Varroa mites infecting their colonies, bear a suite of RNA viruses including major pathogenic viruses like Deformed wing virus, Acute bee paralysis virus, Black queen cell virus and Sacbrood virus, and less common viruses (e.g., bee Macula-like virus and Apis mellifera filamentous virus). The two native honey bee MENA subspecies have acquired different but overlapping suites of pathogens, which also differ, but overlap, with the suites detected in the mites. The presence of plant viruses suggests that they were acquired from foraging for pollen and nectar. Phylogenetic analysis of the above common pathogenic RNA viruses showed unexpected genetic relationships with other known strains, indicative of import to MENA from outside of the region. Our findings indicate that it is important to carefully consider the impact of the movement of queens and mobile colonies, and the effects such movement have, on the transmission of disease.

Description of Parasitic Infections in Honeybees Apis sp in the Middle Euphrates Region of Iraq

The present study was carried out to investigate the parasitic infections of honeybees, 300 adult honey bee collected from apiaries in the Middle Euphrates region (province of AL-Diwaniyah, AL-Najaf, Babylon, Karbala), there were three strains of honeybees in that area included, Apis mellifera syriaca, Apis mellifera syriaca carnica, Apis melliferasyriaca ligustica, all these strains registered were infected with both Parasites Varroa. destructor, Nosema apis where the proportion of total infection was 49.44%, a strain was less parasitic infection Apis melliferasyriaca carnica, it was 8.66% while strain Apis mellifera syriaca more that where 22.66%, the results also showed significant differences in the percentage of infection between the strains of bees at the probability P˂0.05. The results also showed that the incidence of Varroa were 23.33% as single infection, while the prevalence of Nosema was 7.33% as single infection and 18.66% as common infection. The results also indicated that the highest percentage of infection was recorded in the province of Najaf which was 44%, while the lowest percentage of infection was in the province of Diwaniyah, where it reached 31%, and the present study didn't find a relationship between provinces and percentage of parasitic infections in bees.

Prevalence of honey bee (Apis mellifera) parasites across Texas

We conducted a statewide study, in June and July 2016, of feral colonies and domesticated hives to understand the current and potential impacts of invasive parasites and pests on honey bee (Apis mellifera) populations in Texas. We identify the subspecies for each colony sampled in addition to reporting the presence of parasites. The most common subspecies found were Apis mellifera scutellata, Apis mellifera carnica, and Apis mellifera ligustica. We confirmed the presence of Apis mellifera syriaca and Apis mellifera macedonica in Texas. Varroa destructor and Nosema ceranae were the most common parasites, found in eight and nine ecoregions, respectively. Aethina tumida was found in only 4 of the 10 ecoregions.

Effects of different honeybee races on morphological and reproduction characters of honeybee queens grown under Jordan apiaries conditions

This research was conducted during the period of February 2011 to October 2014 at the apiary of the Faculty of Agriculture, Al-Balqa Applied University in Jordan. Honeybee queens of Apis mellifera carnica and Apis mellifera syriaca were raised to investigate some morphological and reproductive characteristics and to determine the most suitable race and hybrid to be reared under Jordan apiaries conditions. Honeybee races and the hybrid between them affected the queens’ weight, pre-oviposition period, volume of spermatheca, number of ovarioles in both ovaries, the length of ovary, beside the lasting of the colony (oviposition period) and the honey production. The hybrid bee race gave the largest queen cell size, the longest queen abdomen, the largest number of ovarioles of both ovaries, and the longest right ovariole, the long lasting colonies with the highest honey production for both the first and second year.

Cover Caption

Honey bees are valuable for agriculture economy and important contributes to ecosystem services due to that they are the most efficient and economic pollinators. However honeybees are facing huge challenges due to degradation of vegetation, and due to honeybee pests and diseases. Apis mellifera syriaca exhibits a high degree of tolerance to pests and pathogens including varroa mites. This native honey bee subspecies of Jordan and most of the Levant region is expresses behavioral adaptations to high temperature and dry seasons typical of the region. Using the next‐generation sequencing (NGS) technology some genetic factors identified which are possibly contributing toward mite resistance and other favorable traits (see pages 579–590). Photo provided by Nizar Haddad.

Evaluation of Apis mellifera syriaca Levant region honeybee conservation using comparative genome hybridization.

Apis mellifera syriaca is the native honeybee subspecies of Jordan and much of the Levant region. It expresses behavioral adaptations to a regional climate with very high temperatures, nectar dearth in summer, attacks of the Oriental wasp and is resistant to Varroa mites. The A. m. syriaca control reference sample (CRS) in this study was originally collected and stored since 2001 from "Wadi Ben Hammad", a remote valley in the southern region of Jordan. Morphometric and mitochondrial DNA markers of these honeybees had shown highest similarity to reference A. m. syriaca samples collected in 1952 by Brother Adam of samples collected from the Middle East. Samples 1-5 were collected from the National Center for Agricultural Research and Extension breeding apiary which was established for the conservation of A. m. syriaca. Our objective was to determine the success of an A. m. syriaca honey bee conservation program using genomic information from an array-based comparative genomic hybridization platform to evaluate genetic similarities to a historic reference collection (CRS). Our results had shown insignificant genomic differences between the current population in the conservation program and the CRS indicated that program is successfully conserving A. m. syriaca. Functional genomic variations were identified which are useful for conservation monitoring and may be useful for breeding programs designed to improve locally adapted strains of A. m. syriaca.

Next generation sequencing of Apis mellifera syriaca identifies genes for Varroa resistance and beneficial bee keeping traits.

Apis mellifera syriaca exhibits a high degree of tolerance to pests and pathogens including varroa mites. This native honey bee subspecies of Jordan expresses behavioral adaptations to high temperature and dry seasons typical of the region. However, persistent honey bee imports of commercial breeder lines are endangering local honey bee population. This study reports the use of next-generation sequencing (NGS) technology to study the A. m. syriaca genome and to identify genetic factors possibly contributing toward mite resistance and other favorable traits. We obtained a total of 46.2 million raw reads by applying the NGS to sequence A. m. syriaca and used extensive bioinformatics approach to identify several candidate genes for Varroa mite resistance, behavioral and immune responses characteristic for these bees. As a part of characterizing the functional regulation of molecular genetic pathway, we have mapped the pathway genes potentially involved using information from Drosophila melanogaster and present possible functional changes implicated in responses to Varroa destructor mite infestation toward this. We performed in-depth functional annotation methods to identify ∼600 candidates that are relevant, genes involved in pathways such as microbial recognition and phagocytosis, peptidoglycan recognition protein family, Gram negative binding protein family, phagocytosis receptors, serpins, Toll signaling pathway, Imd pathway, Tnf, JAK-STAT and MAPK pathway, heamatopioesis and cellular response pathways, antiviral, RNAi pathway, stress factors, etc. were selected. Finally, we have cataloged function-specific polymorphisms between A. mellifera and A. m. syriaca that could give better understanding of varroa mite resistance mechanisms and assist in breeding. We have identified immune related embryonic development (Cactus, Relish, dorsal, Ank2, baz), Varroa hygiene (NorpA2, Zasp, LanA, gasp, impl3) and Varroa resistance (Pug, pcmt, elk, elf3-s10, Dscam2, Dhc64C, gro, futsch) functional variations genes between A. mellifera and A. m. syriaca that could be used to develop an effective molecular tool for bee conservation and breeding programs to improve locally adapted strains such as syriaca and utilize their advantageous traits for the benefit of apiculture industry.

Mitochondrial genome of the Levant Region honeybee, Apis mellifera syriaca (Hymenoptera: Apidae)

The mitochondrial genome sequence of Levant Region honeybee, Apis mellifera syriaca, is analyzed and presented for the public for the first time. The genome of this honeybee is 15,428 bp in its length, containing 13 protein-coding genes, 22 transfer RNA genes and 2 ribosomal RNA genes. The overall base composition is A (42.88%), C (9.97%), G (5.85%), and T (41.3%), the percentage of A and T being higher than that of G and C. Percentage of non-ATGC characters is 0.007. All the genes are encoded on H-strand, except for four subunit genes (ND1, ND4, ND4L, and ND5), two rRNA genes and eight tRNA genes. The publication of the mitochondrial genome sequence will play a vital role in the conservation genetic projects of A. mellifera, in general, and Apis mellifera syriaca, in particular; moreover, it will be useful for further phylogenetic analysis.

Basic considerations in the development of breeding plans for honey bees, illustrated by data on the native Syrian honey bee (Apis mellifera syriaca)

SummaryThe principles of valuing biodiversity are well understood, but methodologies are not yet adequate for enhancing the diversity of honey bee genetic resources. Genetic diversity is generally assumed to be an important barrier against changing environmental conditions, yet some indigenous honey bee breeds are at risk of becoming extinct. The causes of this often stem from development policies in developing countries in the Middle East and other parts of the world, which have largely ignored well-adapted local breeds. The focus instead has been on the introduction of “improved”, exotic ones. Hence, one potential way to ensure survival and promote conservation is by establishing selection programmes that improve efficiency and behaviour in endangered breeds, whilst maintaining their environmental adaptation. However, to develop appropriate genetic models and breeding strategies, genetic aspects and population peculiarities of honey bees must be taken into account. This study aimed at outlining all information needed to plan an efficient honey bee breeding programme, and illustrate this with the example of the endangered Syrian honey bee (Apis mellifera syriaca).

Subjective evaluation of defensive behavior in the Syrian honeybee (Apis mellifera syriaca)

Abstract To separate gentle from defensive bee types, a rating scale is the most reliable, economical, and practical evaluation system. In initiating a breeding programme for the indigenous Syrian honeybee Apis mellifera syriaca, a typical representative of the endangered subspecies of the Middle East, multiple measurements were used to generally monitor this behavior and verify whether the international Apimondia recommendations are suitable for such subspecies. A total of 72 beekeepers and three breeding centers provided performancetesting results from 969 colonies distributed throughout 75 apiaries in Syria. Each colony was tested, on average, 4.73 times (4584 records total). The defensive behavior of A. m. syriaca was found to be very aggressive (1.48 scored by a system ranging from 1 (aggressive) to 4 (gentle)). The low repeatability (0.19) of defensive behavior estimated using restricted maximal likelihood analysis and the low and skewed variation of the trait indicate that other scoring systems are expected to be more efficient for selective breeding towards gentle behavior.

A fifth major genetic group among honeybees revealed in Syria

BackgroundApiculture has been practiced in North Africa and the Middle-East from antiquity. Several thousand years of selective breeding have left a mosaic of Apis mellifera subspecies in the Middle-East, many uniquely adapted and survived to local environmental conditions. In this study we explore the genetic diversity of A. mellifera from Syria (n = 1258), Lebanon (n = 169) and Iraq (n = 35) based on 14 short tandem repeat (STR) loci in the context of reference populations from throughout the Old World (n = 732).ResultsOur data suggest that the Syrian honeybee Apis mellifera syriaca occurs in both Syrian and Lebanese territories, with no significant genetic variability between respective populations from Syria and Lebanon. All studied populations clustered within a new fifth independent nuclear cluster, congruent with an mtDNA Z haplotype identified in a previous study. Syrian honeybee populations are not associated with Oriental lineage O, except for sporadic introgression into some populations close to the Turkish and Iraqi borders. Southern Syrian and Lebanese populations demonstrated high levels of genetic diversity compared to the northern populations.ConclusionThis study revealed the effects of foreign queen importations on Syrian bee populations, especially for the region of Tartus, where extensive introgression of A. m. anatolica and/or A. m. caucasica alleles were identified. The policy of creating genetic conservation centers for the Syrian subspecies should take into consideration the influence of the oriental lineage O from the northern Syrian border and the large population of genetically divergent indigenous honeybees located in southern Syria.