Investigation of genotoxic effects of rhododendron honey using three mammalian bioassays in vivo.

Rhododendron honey (RH) is obtained from the rhododendron plants are grown in many regions around the world, causes poisoning in humans due to the grayanotoxin (GTX) compound in its structure. It is used by the public as a therapeutic for some diseases. It was aimed to study the genotoxic and cytotoxic effects of RH in mouse bone-marrow and sperm cells by using three mammalian bioassays. 25, 50 and 75 mg kg-1 concentrations of RH given to male mice via gavage for 24 and 48 h treatment periods and its active ingredient Grayanatoxin (GTX-III) 0.01 mg kg-1 by i.p. injection. Chromosome aberrations (CA), polychromatic erythrocytes (PCE)/normochromatic erythrocytes (NCE), micronucleated polychromatic erythrocytes (MNPCE) and sperm abnormalities were investigated. The results demonstrated that all the tested concentrations of RH significantly induced total abnormal cell frequency including chromosomal breaks for two time periods. In the MN assay, 75 mg kg-1 RH and 0.01 mg kg-1 GTX-III significantly increased % MNPCE and significantly reduced PCE/NCE ratios after 24 and 48 h treatments on mice demonstrating potential genotoxic and cytotoxic effect. Although there was a concentration-related increase in the percentage of total sperm abnormalities, this increase was not statistically significant compared to control. As a result, microscopic genotoxicity and cytotoxicity marker tests showed that RH and its active ingredient GTX-III have potential genotoxic and cytotoxic effect on mice bone marrow cells. It is understood that RH that is used to treat some diseases by public, should be handled carefully and used in a controlled manner.HighlightsChromosome aberration, micronucleus and sperm morphology assays are recommended as reliable biological indicators.RH and its active ingredient GTX-III have potential genotoxic and cytotoxic effect on mice bone marrow cells.Significant changes were observed upon the treatment of 75 mg kg-1 MH for MN assay.

Authoritative subspecies diagnosis tool for European honey bees based on ancestry informative SNPs.

BACKGROUND: With numerous endemic subspecies representing four of its five evolutionary lineages, Europe holds a large fraction of Apis mellifera genetic diversity. This diversity and the natural distribution range have been altered by anthropogenic factors. The conservation of this natural heritage relies on the availability of accurate tools for subspecies diagnosis. Based on pool-sequence data from 2145 worker bees representing 22 populations sampled across Europe, we employed two highly discriminative approaches (PCA and FST) to select the most informative SNPs for ancestry inference. RESULTS: Using a supervised machine learning (ML) approach and a set of 3896 genotyped individuals, we could show that the 4094 selected single nucleotide polymorphisms (SNPs) provide an accurate prediction of ancestry inference in European honey bees. The best ML model was Linear Support Vector Classifier (Linear SVC) which correctly assigned most individuals to one of the 14 subspecies or different genetic origins with a mean accuracy of 96.2% ± 0.8 SD. A total of 3.8% of test individuals were misclassified, most probably due to limited differentiation between the subspecies caused by close geographical proximity, or human interference of genetic integrity of reference subspecies, or a combination thereof. CONCLUSIONS: The diagnostic tool presented here will contribute to a sustainable conservation and support breeding activities in order to preserve the genetic heritage of European honey bees.

Comparing the biological properties and chemical profiling of chestnut bee pollen and bee bread collected from Anatolia.

Bee bread (BB) and bee pollen (BP) are accepted as functional food and considered in medical properties due to its important bioactive components. These bee products show different biological properties, but researches on these aspects have not been clear yet. In present study, Anatolian BB and BP extracts were analyzed for the first time for their pollen type, total phenolic (TPC) and flavonoid content (TFC), and antimicrobial and antioxidant properties. Samples were analyzed for their antimicrobial efficacy by the agar well diffusion and MIC methods. HPLC analysis was used to identify the compounds in the BB and BP samples. Antioxidant activity was measured by the FRAP and DPPH methods. As a result of microscopy for pollen identification, Fagaceae family was dominant. Phenolic compound analysis showed that the amounts of p-coumaric acid and rutin were found to be the highest in BB and BP, respectively. Stronger antioxidant activity was obtained from BP. MIC values of BB were range from 250 to 12.5 µg/mL. The most susceptible bacterium was Mycobacterium smegmatis. The extract of BP was effective on all gram-negative bacteria with doses range from 250 µg/mL to 500 µg/mL. The lowest MIC value was detected with the concentration of 12.5 µg/mL against M. smegmatis. Anatolian BB and BP could be considered as a functional foods due to antioxidant activity and may be beneficial in the management and treatment of pathogenic bacteria because of high antimicrobial activity.

Chemical profiling and antimicrobial effect of Anatolian honey bee venom.

Due to increasing antibiotic resistance, there is an urgent need to find new antibiotic alternatives or supporters for the treatment of disease-causing pathogens. For this reason the aim of the study was examine the antimicrobial and antifungal activity of Anatolian (Anadolu) honey bee venom (HBV) against Gram-positive and Gram-negative bacteria and yeast-like fungi. At first step chemical analyses of HBV was performed by HPLC method. According to the results of HPLC analysis, we obtained a good separation of apamine, phospholipase A2 and melittin with the ratio of 1.83%, 20.60% and 57.62% respectively. The antimicrobial and antifungal activity of the Anatolian HBV was tested against 9 Gram (+), 7 Gram (-), 1 acid-alcohol-resistant and 3 yeast fungi. First, the activity of the Anatolian HBV sample against these microorganisms was determined by the agar well diffusion method, then their zones were measured. The microdilution method was used to determine the minimum inhibitory concentration (MIC) for the antimicrobial activity tests. The results of MIC values were varied from 3.06 µg/mL to 50 µg/mL for the tested microorganisms. It was found that Mycobacterium smegmatis and Streptococcus pyogenes were the most susceptible bacteria (3.06 µg/mL), followed by Vibrio sp., Aeromonas sobria, Enterococcus faecalis, Listeria monocytogenes, Staphylococcus aureus (MRSA) and B. subtilis with a MIC concentration of 6.125 µg/mL. These findings strongly suggest that Anatolian HBV will be developed as a new antibacterial-antifungal drug against Gram-positive, Gram negative and antibiotic-resistant bacteria and yeast-like fungi. However, further research is required to evaluate their in vivo efficacy and safe and effective delivery methods for their therapeutic use.

Development of a cream formulation containing bee venom and other bee products.

This study aimed to develop a prototype skincare product with bee venom, propolis, honey, beeswax, and royal jelly. The prototype formulation contained 0.1% bee venom, 0.3% propolis extract, 0.45% honey, and 1.0% royal jelly. The prototype body cream was analyzed for stability, antioxidant activity, dermatological response, and cytotoxicity. In addition, a panel test evaluated the prototype for the claims such as skin smoothness, feelings of nourishment, moisturizing, skin tone, brightness, and visibility of wrinkles. According to the stability test, the prototype was stable for up to 90 days at room temperature and +40°C. The formulation was found to have a high antioxidant capacity at 85.45%. Cell viability detected over 70% indicated that the prototype body cream was not cytotoxic. The dermatological analysis revealed no irritation or allergic reaction in non-allergic individuals. Panel test showed that the prototype makes skin silky smooth, contributes to hydration, brightens and nourishes the skin, evens the skin tone, reduces the visibility of wrinkles, improves skin elasticity, and smoothes wrinkles. This prototype formulation requires further research to evaluate its effectiveness against skin aging on different skin types. Nevertheless, the side effects of such products need particular attention in developing a commercial product containing bee venom in susceptible individuals.

Protective effect of propolis on myocardial ischemia/reperfusion injury in males and ovariectomized females but not in intact females.

The aim of this study is to investigate the effect of propolis, which may have estrogenic effects, on myocardial ischemia/reperfusion (mI/R) injury not only in male rats but also in intact and ovariectomized (ovx) female rats. Six groups were formed: untreated males (n = 8), treated males (n = 9), untreated intact females (n = 9), treated intact females (n = 10), untreated ovx females (n = 10), and treated ovx females (n = 8). An alcoholic extract of a single dose of propolis (200 mg/kg) was administered orally daily for 14 days. Thirty minutes of ischemia and 120 min of reperfusion were performed. Blood pressure, heart rate, arrhythmias (ventricular premature contraction [VPC], ventricular tachycardia [VT], ventricular fibrillation [VF]), and myocardial infarct size were evaluated. Total antioxidant status (TAS), total oxidant status (TOS), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and 17 beta-estradiol (E2) were measured. The untreated females showed more resistance to mI/R injury than the untreated males, as evidenced by lower duration, incidence, and score of arrhythmias, and smaller infarct size (p < .05). After ovx, this resistance disappeared. Propolis improved these values in treated males and treated ovx females (p < .05). Propolis increased TAS in treated males and decreased TOS in treated ovx females as well as elevated SOD in all treated groups (p < .05). Propolis decreased E2 level in treated intact females; however, it increased E2 level in treated ovx females (p < .05). The results revealed that propolis could protect the heart against mI/R injury in males and ovx females. PRACTICAL APPLICATIONS: It is known that the female heart has an increased sensitivity to myocardial ischemia/reperfusion (mI/R) injury due to estrogen deficiency and/or estrogen deprivation following menopause or surgical removal of the ovaries. Propolis has the potential to mimic estrogen under physiological and pathophysiological conditions, as well as its antioxidant property. The results indicated that propolis decreased myocardial infarct size, arrhythmia score, arrhythmia duration, and incidence in ovariectomized female rats and male rats. In addition, the present results demonstrated that an alcoholic extract of propolis as a natural product can effectively maintain the resistance of female heart to mI/R injury after estrogen deficiency.

An evaluation of the chemical content and microbiological contamination of Anatolian bee venom.

Bee venom is a natural substance produced by worker bees. The aim of this research paper is to determine the characteristics of Anatolian bee venom by evaluating its chemical content and microbiological properties. Physical, chemical and microbiological analyses were performed on 25 bee venom samples from different areas of Anatolia, Turkey. Data obtained by 3-replicate studies were evaluated with normality and one-way and two-way ANOVA / Tukey tests. Chemical analyses of the bee venoms revealed average melittin, apamin, and phospholipase A2 contents of 40.57%, 2.12% and 13.67%, respectively. The results suggest that Anatolian bee venom has a high phospholipase A2 content compared to the previous literature. The results for apamin content were similar to those reported in other countries. Melittin content was within the range of standard values. Bee venom samples were also observed to have a high sugar content, associated with pollen and nectar contamination. Total aerobic mesophilic bacteria counts revealed no microbial development in 11 samples of bee venom. Staphylococcus aureus was not detected in any sample. A low microbial load was associated with a high phospholipase A2 content in the bee venom composition, thus contributing to its antimicrobial character. This study presents an examination of Anatolian bee venom in terms of chemical content and microbial quality. The examination of other components in addition to phospholipase A2, melittin and apamin in future studies, together with an analysis of antimicrobial properties will further our understanding of Anatolian bee venom.

Determination of the dose-dependent toxic effects of mad honey on mouse liver using ATR-FTIR spectroscopy.

Mad honey (MH) is obtained from Rhododendron plants, which are extensively grown in some regions of the world such as Europe, North America, Tropical Asia and Turkey. Although it has been known that MH induces adverse effects in the body due to grayanotoxin (GTX) in it, it is widely used for some medical purposes by the public. In this study, the effects of MH (25, 50 and 75 mg/kg) and GTX-III (0.01 mg/kg), which is the pure form of the most toxic type of the GTXs in MH, were investigated on the mouse liver at molecular level via Attenuated Total Reflection-Fourier Transform Infrared (ATR-FTIR) spectroscopy. The results showed that 25 and 50 mg/kg of MH didn't cause any significant alterations in the liver tissue except a decrease in the glycogen amount. However, significant differences were observed between 75 mg/kg MH and GTX-III treated groups and control group. For example, the amounts of saturated lipids, nucleic acids and proteins increased in the 75 mg/kg MH and GTX-III treated groups. A decrease in the ratios of unsaturated/saturated lipid, CH2/lipid and carbonyl/lipid and an increase in the ratio of CH3/lipid were observed after the administration of 75 mg/kg MH and GTX-III, all of which may be a consequence of lipid peroxidation. Moreover, 75 mg/kg MH and GTX-III caused a decrease in the membrane order, an increase in the membrane fluidity and some important changes on the secondary structure of proteins indicating protein denaturation. In addition, Hierarchical Cluster Analysis (HCA) and Principal Component Analysis (PCA) confirmed these findings. These results revealed that MH induces significant dose-dependent toxic effects in the structure and function of the liver tissue. This study also showed that ATR-FTIR spectroscopy provides a rapid and sensitive monitoring of the changes induced by a toxic compound on biological tissues at molecular level.

The effect of rhododendron honey on mice liver tissue / Efecto de la miel de rododendro en el tejido hepático de ratón

Rhododendron honey, made by bees from rhododendron pollen, contains a toxic substance called grayanotoxin. Depending on the dose, the poisonous honey can result in serious effects such as cardiac arrhythmia, fibrillation, and myocardial infarction. The purpose of this study is to investigate the effects of the poisonous RH of the Black Sea Region on the liver. Male mice were divided into five groups of twelve mice each, two being the control groups (distilled water) and the others being the rhododendron honey (RH) groups (25, 50, and 75 mg/kg) and 0.01 mg/kg grayanotoxin (GTx) groups. Liver tissues were collected 24 and 48 h later. The sections were stained with hematoxylin, eosin and PAS, then the histopathological score was performed. Significant statistical differences were observed between the RH and control groups in terms of congestion, steatosis, sinusoid dilatation, and inflammation. The control group demonstrated a normal liver structure in the light microscopy, while the GTx-applied 24 h group exhibited expansions in the sinusoids and congestion. Higher levels of congestion, steatosis, and inflammatory cells were seen in the GTx-applied 48 h group. In the same group, giant cells consisting of many nuclei were observed in the sinusoids. The results of the 25 mg RH-applied groups were similar in 24 and 48 h, histopathological score levels were increased slightly, congestion and steatosis were prominent in the 48 h group. Dense steatosis was seen in the hepatocytes around the vena centralis in 50 mg/kg RH-applied 48 h group. Congestion, steatosis and an increase in inflammatory cells were observed in the hepatocytes in the 75 mg/kg RH-applied 24- and 48 h groups. PAS (+) stained hepatocytes were decreased in the RH- and GTx-applied groups. The toxic effects of the rhododendron honey were observed in the mice liver tissue with respect to dose and time.

La miel de rododendro, elaborada por las abejas a partir del polen de rododendro, contiene una sustancia tóxica llamada grayanotoxina. Dependiendo de la dosis, la miel venenosa puede resultar en efectos graves, tales como arritmia cardiaca, fibrilación e infarto de miocardio. El propósito de este estudio fue investigar los efectos en el hígado de la miel venenosa de rododendro de la región del Mar Negro. Se distribuyeron ratones machos en cinco grupos de doce ratones cada uno, dos grupos control (agua destilada) y los otros grupos se trataron con la miel de rododendro (MR) (25, 50 y 75 mg/kg) y con 0,01 mg/kg grayanotoxina (GTX). Los tejidos hepáticos se recogieron 24 y 48 h más tarde. Las secciones fueron teñidas con hematoxilina-eosina y PAS. A continuación, se realizó la puntuación histopatológica. No se observaron diferencias estadísticamente significativas entre MR y los grupos de control en términos de congestión, esteatosis, dilatación sinusoidal e inflamación. El grupo control demostró una estructura normal del hígado en el microscopio de luz, mientras que el grupo de las 24 horas de aplicación de GTX exhibió expansiones en los sinusoides y congestión. Mayores niveles de congestión, esteatosis y células inflamatorias se observaron en el grupo de 48-horas de aplicación de GTX. En el mismo grupo, se observaron células gigantes que consistían en la presencia de muchos núcleos en los sinusoides. Los resultados de los grupos con aplicación de 25 mg de RH fueron similares en los resultados de 24 y 48 h, los niveles de puntuación histopatológica aumentaron ligeramente, la congestión y la esteatosis fueron prominentes en el grupo de 48 h. Se observó esteatosis densa en los hepatocitos en toda la vena central en el grupo de aplicación de 50 mg/kg de RH, 48 h. La congestión, la esteatosis y un aumento en las células inflamatorias se observaron en los hepatocitos en el grupo de 75 mg/kg de MR de 24 h y los grupos de 48 h. Hepatocitos teñidos con PAS (+) disminuyeron en los grupos de GTX y MR. Se observaron los efectos tóxicos de la miel de rododendro en el tejido hepático de ratones con respecto a la dosis y el tiempo.