Pine Honey Research Articles

The effect of pine honey on the viability of probiotics and some properties of probiotic yogurt

In this study, it was aimed to investigate the effect of pine honey on the viability of probiotic bacteria (Lactobacillus acidophilus LA-5®️ and Bifidobacterium animalis subsp. lactis BB-12®️) and some properties of probiotic yogurts. The non-fat dry matter of milk used in yogurt production is standardized to 11%. Milk is divided into 3 groups (A, B, and C). A, B, and C milk groups were inoculated with yogurt culture, yogurt culture + L. acidophilus LA-5 and yogurt culture + B. animalis subsp. lactis BB-12 respectively. Each 3 groups of milk are divided into 3 parts. While pine honey was not added to the first part (control group), 5% pine honey was added to the second part and 7% pine honey to the third part of the milk. Produced yogurt samples were stored at +4 °C for 21 days and physico-chemical, structural, microbiological, and sensory analyzes were performed on the 1st, 7th, 14th and 21st days of storage. According to the results obtained in the study, with the addition of pine honey, hardness and viscosity values of yogurt samples decreased, while water holding capacity values increased. Also, pine honey addition did not have a significant effect on L. acidophilus LA-5 viability, however, the addition of 7% pine honey statistically significantly increased the viability of B. animalis subsp. lactis BB-12. Also, taste scores of yogurts containing probiotic cultures are lower than those of the control group yogurt in sensory analysis. It was determined that the odor, consistency and color scores of the samples were close to each other. Keywords: Pine Honey; yogurt; prebiotic; probiotic; viscosity

QUALITY EVALUATION OF PINE AND BLOSSOM HONEY SAMPLES PRODUCED IN TURKEY: CORRELATION BETWEEN PHYSICOCHEMICAL CHARACTERISTICS

This study presents physicochemical characteristics of 39 honey samples (21 blossom and 18 pine honey) collected during two years from three different geographical regions of Türkiye that differs vastly in climatic conditions and thus plant species. The samples were analysed for δ13C/δ12C stable carbon isotope ratios of honey (δ13Ch) and its protein fraction (δ13Cp), moisture, free acidity, proline and 5-hydroxymethyl furfural (HMF) content, diastase activity and sugar composition. The results showed that C4 sugar content, proline content, diastase activity, acidity values of pine honeys were higher than that of blossom honeys whereas, higher moisture and HMF content were detected for blossom honeys. Besides, geographical region mainly affected the moisture and C4 sugar contents. High correlations between HMF and δ13Ch and δ13Cp; proline and acidity values; fructose and glucose content were determined, and this indicated the robustness of the analysis and quality evaluation among different honey types and regions.

Determination of forest fire risk with respect to Marchalina hellenica potential distribution to protect pine honey production sites in Turkey.

Turkey is the leading producer of pine honey worldwide, accounting for 90% of global production, largely due to the presence of Marchalina hellenica populations. However, in recent years, devastating forest fires have caused substantial damage to Pinus brutia forests and M. hellenica populations, leading to a dramatic decline in pine honey production areas. The urgency for early intervention procedures against forest fires and relocation of M. hellenica populations to other P. brutia forests has become apparent. A comprehensive assessment of 25 criteria was conducted to investigate the thresholds and behaviors of each criterion, which play a vital role in the distribution of M. hellenica, using the maximum entropy model (MaxEnt). To evaluate the impact of forest fires on the distribution of M. hellenica, the potential locations of pine honey production areas were determined for 2022. Furthermore, the susceptibility of forest fires was modeled for all pine honey production months. The findings revealed that forest fires have destroyed 384.9 km2 (12.8% of the total pine honey production area), predominantly in August and September, with the most severe damage in Marmaris (156 km2) and significant impacts in Ula, Köyceğiz, and Milas. The analysis facilitates the estimation of new areas suitable for M. hellenica and pine honey production while providing valuable insights into strategies for mitigating forest fires and formulating proactive protection protocols.

Distinguishing Turkish pine honey from multi-floral honey through MALDI-MS-based N-glycomics and machine learning

Honey, a multifaceted blend of sugars, amino acids, vitamins, proteins, and minerals, exhibits compositional variability dependent upon the floral source. While previous studies have attempted to categorize honey, the use of glycomic profiles for honey classification remains an unexplored avenue. This investigation seeks to establish a methodology for distinguishing honey types, specifically multi-floral and pine honey, employing mass spectrometry-based glycomic analysis in tandem with machine learning. In this search, seven samples of pine honey and eight samples of multi-floral honey were obtained from diverse regions of Turkey. Subsequently, the proteins within these honey samples were extracted, and glycans were enzymatically released. The released glycans were labeled with 2-aminobenzoic acid (2-AA) and subjected to analysis via matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The glycan profiles of pine and multi-floral honey were determined through these analytical procedures, revealing a total of 76 distinct N-glycan structures. Among these, 13 N-glycan profiles consistently established at high levels across experimental replicates and were incorporated in subsequent analyses. Following the quantification of individual glycan abundances, statistically significant differences in glycan profiles were determined. Notably, N-glycans Hex5HexNAc2, Hex4HexNAc3, and Hex5HexNAc3 displayed considerable differences. Using the 13 N-glycan profiles, an accuracy rate of 93.5% was obtained from machine learning analysis, which increased to 100% when incorporating the identified significantly changed glycans. The most productive models were identified as “subspace and fine k-nearest neighbors (KNN).” The findings underscore the potential of mass spectrometry-based glycomics in conjunction with machine learning as a robust tool for precise honey type classification and its prospective utility in quality control and honey product authentication.

Determination of intestinal microbiome of Marchalina hellenica (Gennadius, 1883) (Hemiptera, Margarodidae)

Knowledge of the insect microbiota remains largely unexplored. The intestinal microbiome of Marchalina hellenica (Gennadius, 1883) (Hemiptera, Margarodidae) was determined in the current study. This species is the main source of pine honey production. A pool of 100 samples were selected, representing the distribution of M. hellenica in the province of Mugla, Turkey. API Staph tests were used to assess Gram-positive bacteria, while API 20NE and API 20E tests were used to assess Gram-negative bacteria. As a result, 33 Gram-negative and 15 Gram-positive isolates were obtained from samples collected from 44 different locations. Among Gram-positive, 14 out of 15 belonged to Staphylococcus spp. Among Gram-negative, 22 out of 33 belonged to Aeromonas spp. This is the first study intended to determine the intestinal microbiome of M. hellenica.

Characterization of Turkish Pine honey and differentiation from floral honeys by NMR spectroscopy and chemometric analysis

Honey is a viscous, supersaturated sugar solution produced by bees through the enzymatic transformation of nectar from flowers, containing a complex mixture of carbohydrates, organic acids, enzymes, and other minor constituents. Although honey has been used for thousands of years for its nutritional and medicinal properties, it has been the subject of increasing attention in recent years due to increasing adulterated honey production. Consequently, assessment of honey quality and authenticity has become essential to ensure consumer confidence of regional honey and to perverse the practice of authentic honey production. In this study, we employed nuclear magnetic resonance (NMR) spectroscopy and chemometric analysis to characterize Turkish Pine honey and compare it to flower honey originating from the Oceania, Aegean, and West Coast of North America regions. Utilizing 1H NMR spectroscopy, the chemical profile of Turkish Pine honey was characterized, and unique peaks were found. Additionally, PLS-DA statistical analysis was employed to further investigate the distinction of Turkish Pine honey among other floral and regional honeys. Upon completion of our statistical analysis, we were able to effectively distinguish Turkish Pine honey from other regional honey types, allowing us to formulate a universal test for authenticity of Turkish Pine Honey.

Effects of the addition of flower honey and pine honey to extenders on spermatological characteristics in ram semen

Bu çalışmada, belirli oranlarda çiçek ve çam balının koç spermatının spermatolojik parametreleri üzerine etkileri araştırılmıştır. Tris-yumurta sarısı bazlı meni sulandırıcıya her bal türü için %1.0 (h/h) ve %2.5 (h/h) oranlarında çiçek ve çam balı eklenmiştir. Sperm toplama için beş Kıvircik koç kullanıldı. Toplanan spermler havuzlamanın ardından 5 eşit parçaya bölündü. %1.0 (h/h) ve %2.5 (h/v) çam balı ve %1.0 (h/v) ve %2.5 (h/h) çiçek balı oranlarında ve bal ilavesi yapılmadan iki aşamalı seyreltme yöntemi kullanılmıştır. Tris-yumurta sarısı bazlı meni genişleticiye. Numuneler daha sonra sıvı nitrojen buharında dondu ve sıvı nitrojen içinde saklandı. Semen canlılığı parametreleri (toplam ve progresif motilite, hız, kinetik hareket parametreleri ve mitokondriyal aktivite) ve morfolojik bütünlük parametreleri (Hancock fiksasyon testi, akış sitometrisi ve floresan boyama) dengeleme ve çözdürmeden sonra değerlendirildi. Çalışma boyunca on tekrarlı çalışma yapılmıştır.Sonuçlar çiçek balının çözülme sonrası canlılık parametreleri üzerinde özellikle %2,5 (h/h) oranında faydalı etkiye sahip olduğunu göstermiştir. Her oranda ve her çeşit bal içeren sulandırıcılar, çözüldükten sonra zar bütünlüğünü korumuştur. Tris-yumurta sarısı bazlı sulandırıcıya %1.0 (v/v) çiçek balı ilavesi, diğer gruplara kıyasla Hancock fiksasyon testi ile akrozomal bütünlük açısından daha iyi sonuçlar vermiştir (p < 0.05). Sonuç olarak, Tris-yumurta sarısı sulandırıcısına %1.0 (h/h) ve %2.5 (h/v) çiçek balı ilavesiyle çözülme sonrası spermatolojik parametrelerde daha iyi sonuçlar kaydedildi.

Pesticide residues in different honey types and public health risk assessment

Honeybees and humans are endangered by pesticides in daily agricultural production. The aim of this research was to investigate pesticide residues in different honey types and to assess the risk to public health. A total of 88 honey samples originating from pine, multifloral, sunflower, acacia, linden, and canola were collected and analysed by a QuEChERS method. The hazard quotient (HQ) was used to evaluate the risk of detected pesticide residues. Analysis of pine honey did not detect any residue of investigated pesticides. The most frequently detected pesticides in the honey samples were chlorpyrifos ranging between 15.1 µg/kg (linden honey) to 22.3 µg/kg (multifloral honey), clothianidin ranging between 12.0 µg/kg (acacia honey) to 22.0 µg/kg (canola honey), dimethoate ranging between 8.9 µg/kg (multifloral honey) to 18.9 µg/kg (canola honey), and thiamethoxam ranging between 4.2 µg/kg (linden honey) to 15.6 µg/kg (canola honey), respectively. The lowest estimated daily intake (EDI) of 128 × 10-3 μg/kg of body weight per day was found in acacia honey, and the highest EDI of 265 × 10-3 μg/kg of body weight per day was found in canola honey. Similar values of EDI were determined for multifloral, sunflower, and linden honey (186 × 10-3, 187 × 10-3, and 183 × 10-3), respectively. The HQ value for pine honey was 0 indicating that this honey is the safest for consumption, however, the other types of honey investigated in this study posed no risk to humans after potential consumption.

Development and validation of a high-throughput headspace solid-phase microextraction gas chromatography-mass spectrometry methodology for target and suspect determination of honey volatiles

The determination of volatile compounds is essential for the chemical characterisation of honey's aroma and its correlation to its sensory profile and botanical origin. The present study describes the development, optimization and validation of a new, simple and reliable method for the determination of volatile compounds in honey using headspace solid-phase microextraction combined with gas chromatography/mass spectrometry (HS-SPME-GC-MS). The optimization of the SPME conditions showed that the ratio of honey: water (2:1) and the incubation temperature (60 °C) are the most critical parameters. Gas chromatography was performed with medium polar Varian CP-Select 624 column and the experimental Retention Index for a number of compounds was determined as an additional identification feature for suspect analysis. The simultaneous use of four internal standards chlorobenzene, benzophenone, 2-pentanol and 4-methyl-2-pentanone and matrix matched calibration enhanced method accuracy achieving recoveries 73–114 % and repeatability ranging between 3.9 and 19 % relative standard deviations. Furthermore, the superiority of the HS-SPME to static head space technique was verified exhibiting four-to nine-fold higher sensitivity. Target and suspect screening were applied to 30 Greek honey samples and 53 volatile compounds belonging to different chemical classes, such as alkanes, aldehydes, ketones, alcohols, and esters were identified with quantified concentrations ranging between 3.1 μg kg−1 (Limonene) up to 20 mg kg−1 (Benzeneacetaldehyde). Among the new findings is the detection of Myrtenol in Greek pine honey and 2,3-butanediol in Greek oak honey. The developed analytical protocol can be a valuable tool in order to chemically characterize honey based on the volatile content.

Effect of different fermentation conditions on bioactive properties, phenolic component and sensory properties of caper (Capparis ovata desf. var. ovata) buds

In this study, the antioxidant activities of caper buds, total phenolic acid content, carotenoid, flavonoid, phenolic components and total phenolic acid content of caper buds after fermentation of caper flower buds in separately 10% brine (NaCl2) containing 5% flower honey, 5% pine honey, 5% granulated sugar and 5% glucose syrup and to reveal their sensory properties were investigated. The difference of this study from other studies is the use of honey, glucose and saccharide (sugar) in addition to the use of salt in lactic acid fermentation, the main difference being the use of caper buds and the innovation. Total carotenoid contents of raw (control) and fermented caper buds were determined between 0.27 µg/g (Glucose supplemented) and 9.40 µg/g (control (raw)). In addition, the total phenolic content of raw and fermented caper buds was measured between 106.45 mg gallic acid equivalent (GAE)/100 g (raw) and 231.07 mg GAE/100 g (with added sugar), while the total flavonoid values of raw and fermented caper buds are determined between 242.26 mg quercetin equivalent (QE)/100 g (raw) to 453.48 mg QE/100 g (sugar added). In addition, antioxidant activity values of raw and fermented caper buds were measured between 6.04 mmol trolox equivalent (TE)/kg (with added salt) and 7.75 mmol TE/kg (with added sugar). Gallic acid, 3,4-dihydroxybenzoic acid and catechin were the dominant phenolic components of raw and fermented caper buds. Caper buds fermented in brine with added sugar and glucose were the most appreciated.

Characterization of Turkish pine honey according to their geographical origin based on physicochemical parameters and chemometrics

This study was conducted to determine the characteristic properties of Turkish pine honey, which is an important honeydew honey. The geographical classification of the honey was determined by applying carbon isotope, melissopalynological, and physicochemical analyses to 373 samples collected from 47 regions between 2015 and 2017 under controlled conditions. δ13C protein-δ13Choney, C4%, electrical conductivity, moisture, ash, free acidity, color CIEL* a*b attributes, optical rotation [α]20, proline, diastases activities, and sugars (fructose, glucose, sucrose, and maltose) were used as physicochemical properties. Number of honeydew elements /number of total pollen (NHE/NTP) ratios were studied at melissopalynological analyses. The results showed that all samples exhibited honeydew properties, and that all physicochemical parameters met the criteria set by regulatory standards for honeydew. However, C4% sugar and δ13C protein-δ13C honey values did not meet the regulatory criteria and exhibited quite wide standard deviations.

Solid-phase microextraction Arrow combined with comprehensive two-dimensional gas chromatography-mass spectrometry for the elucidation of the volatile composition of honey samples.

In this work, a solid-phase microextraction (SPME) Arrow method combined with comprehensive two-dimensional gas chromatography-mass spectrometry (GC × GC-MS) was developed for the elucidation of the volatile composition of honey samples. The sample preparation protocol was optimized to ensure high extraction efficiency of the volatile organic compounds (VOCs) which are directly associated with the organoleptic properties of honey and its acceptance by the consumers. Following its optimization, SPME Arrow was compared to conventional SPME in terms of sensitivity, precision, and number of extracted VOCs. The utilization of SPME Arrow fibers enabled the determination of 203, 147, and 149 compounds in honeydew honey, flower honey, and pine honey, respectively, while a significantly lower number of compounds (124, 94, and 111 for honeydew honey, flower honey, and pine honey, respectively) was determined using conventional SPME. At the same time, the utilization of SPME Arrow resulted in enhanced sensitivity and precision. All things considered, SPME Arrow and GC × GC-MS can be considered as highly suitable for the elucidation of the volatile composition of complex food samples resulting in high sensitivity and separation efficiency.

Static Water Vapor Sorption Properties of Honey

Abstract Static sorption properties of pine honey (PH) and citrus honey (CH) were studied at 20, 30 and 40°C. Yeast formation was observed on the surface of honeys at water activity (aw)>0.7 at all temperatures. Visible yeast formation (YF) took place earlier in PH than in CH under the same conditions due to its characteristic higher pH and lower monosacchride content. The temperature was insignificantly effective on YF (p>0.05). The honeys exhibited a sorption isotherm (SI) in the shape of a “J”. Their SIs exhibited desorption and adsorption at aw<0.7 and aw>0.7, respectively and the desorption part was almost linear. Aw=0.7 emerged as a border between the absence and presence of YF and between the desorption and adsorption. The SIs of honeys were insignificantly affected by temperature (p>0.05). PH had a significantly lower SI than that of CH (p≤0.05) due to its characteristic lower monosacchride content. GAB equation exhibited a good fit to the honeys' SIs. Sorption heat vs equilibrium moisture content (EMC) revealed monolayer, multilayer and loosely bound free water regions in the SIs of PH and CH.

Identification of the Main Phenolic Markers in Turkish Pine Honeys and Their Biological Functions.

The purpose of this research was to identify the biological activity of Turkish pine honeys and the most abundant phenolic compound(s) to guide future studies to be done about this type of honey. Moreover, this will provide the chance to introduce Turkish pine honeys by elucidating a distinct parameter compared to other forms of honey reported in the literature. To this end, microscopic examination based on the number of honeydew elements (NHE) and the number of total pollen (NTP) grains was performed on 17 samples of pine honeys from the Aegean region of Turkey. Then 18 commonly-occurring phenolic standards were used in high-performance liquid chromatography (HPLC) to examine the samples. Each sample was tested for individual phenolic compounds, as well as total phenolic content (TPC), total flavonoid content (TFC), and ferric reducing antioxidant power (FRAP). As shown by the numerical data, the values for NHE and NTP were 6600-144000 and 3000-75000, respectively. In addition, the general means of TPC, TFC, and FRAP values for the samples were 53.000 mg GAE/100 g fresh sample, 1.944 mg QE/100 g fresh sample, and 3.455 μmol FeSO4 /g fresh sample, respectively. Protocatechuic acid predominated with a range of 197.841-780.227 μg phenolic /100 g sample in all samples. According to the general mean for values detected in the samples, p-hydroxybenzoic acid and benzoic acid were the second and third markers for Turkish pine honey samples. It is believed that these results will help efforts to set standards for pine honey on a national and global scale.

The Effect of Selected Bee Products on Adhesion and Biofilm of Clostridioides difficile Strains Belonging to Different Ribotypes.

There is an ongoing search for alternative treatments for Clostridioides difficile infections. The aim of the study was to investigate the antibacterial and antibiotic activity of bee products against C. difficile strains with different polymerase chain reaction ribotypes (RTs). The minimum inhibitory concentration (MICs) of Manuka honey 550+, goldenrod honey, pine honey, and bee bread were determined by the broth dilution method. C. difficile adhesion to HT-29, HT-29 MTX, and CCD 841 CoN cell lines was assessed. Biofilm was cultured in titration plates and visualized by confocal microscopy. The MICs of Manuka honey for C. difficile 630 and ATCC 9689 strains and control strain, M 120, were 6.25%, 6.25%, and 1.56% (v/v), respectively; of goldenrod honey, 50%, 50%, and 12.5%, respectively; of pine honey, 25%, 25%, and 25%, respectively; and of bee bread, 100 mg/L, 50 mg/L, and 100 mg/L, respectively. Manuka honey (1%) increased adhesion of C. difficile RT176 strains, and one strain of RT023, to the CCD 841 cell line. Pine honey (1%) increased RT027 adhesion to the HT-29 cell line. Manuka honey, pine honey, and bee bread at subinhibitory concentrations increased the adhesion of C. difficile. Our research proved that bee products are active against the tested strains of C. difficile.

Natural Disaster Risk Assessments for Pine Honey Apiaries in Muğla, Turkey

Since Muğla province has 90% of the world's total pine honey production, ensuring efficiency and economic income requires the determination of measures for apiary locations and estimation of risks. However, ensuring development and productivity requires identifying natural disasters susceptibility such as forest fires and floods to maintain productivity. Muğla province has a high forest fire potential due to its dense forest cover and approximately 200 forest fires occur each year. Forest fires are one of the main factors that threaten apiaries, as there are a lot of apiary places (approximately 15,000) in forests for pine honey. On the other hand, due to the mountainous topography and high precipitation rate of Muğla, the province has a high rate of flood formation (20 per year), which threatens the hive sites by destroying the entire colony. In this study, Apiary Locations Risk Index (ALRI) was carried out to guide the insurance process for apiary locations by applying the Forest Fire Risk Index (FFRI) and the Flood Hazard Risk Index (FHRI). Determination of forest fire risk zones and flood hazard maps requires environmental, forestry, topographic, economic and meteorological parameters to be handled within a decision support platform. For this purpose, Analytical Hierarchy Process (AHP) technique supported by Geographic Information System (GIS) was used in the creation of sensitivity maps. As a result, 1533.40 ha (11.82%) of the study area was determined as extremely risky areas for apiary areas. The results were confirmed with 1454 forest fire sites and 20 flood hazard sites where the Eşen, Dalaman, Çine, Sarıçay, Akçay, Kamiişdere and Namnam rivers were stated to be highly susceptible to flood hazard.

Elucidating the false positive tendency at AOAC 998.12 C-4 sugar test for pine honey samples: Modified sample preparation method for accurate δ13C measurement of honey proteome

The majority of the authentic pine honey samples fail from C-4% sugar test. Pine honey is generally characterized by small-sized insoluble honeydew elements, and these can form an appreciable amount of precipitate during protein flocculation. These substances may shift the protein-centric δ13C (13C/12C) values inaccurately. In this study, honey proteins were isolated, enriched, and cleaned-up prior to flocculation by applying the optimized and validated ultrafiltration method. Authentic honeys were analyzed along with adulterated samples using both AOAC 998.12 and novel protocol. When the interfering substances were eliminated, most of the samples, which were interpreted as adulterated according to the AOAC's method previously, were identified as authentic. It has been proven that inconsistent protein measurements may cause false positivity. These findings revealed that the developed method could correct the artificially high C-4% results and the proposed modification will pave the way for increasing the overall reliability of the test.

Investigation of the effects of Pine and Chestnut Honey on wound healing

This study aimed to investigate the effect of creams containing pine and chestnut honey on wound healing in rats. The animals were divided into the following four groups: a control group (A), a group treated with only basic cream ingredients (B), a group treated with pine honey cream (C), and a group treated with chestnut honey cream (D). Full-thickness skin wounds were created on the back of each rat (14 per group) with a 10-mm diameter punch instrument. In each group, skin samples were taken from seven rats on day 7 and from the remaining seven rats on day 14. The effects of the creams on wound healing were assessed by histopathological examination and hydroxyproline assays. The histopathological examination showed that chronic inflammation significantly decreased in group D compared to the control group (P<0.05). Furthermore, the levels of granulation tissue and granulation tissue/fibroblast maturation in skin samples taken on day 14 were also significantly lower in group D compared to the control group (P<0.05). It was observed that the neovascularization values of skin samples taken on day 7 were lower in group D than in the control group (P<0.05). Also, the difference in hydroxyproline levels between groups B and D was statistically significant (P<0.05). The results showed that the cream containing chestnut honey had a positive effect on wound healing.

The Effect of Supplementary Feeding with Different Pollens in Autumn on Colony Development under Natural Environment and In Vitro Lifespan of Honey Bees.

Simple SummaryIn the present study, the effect of feeding with pollen sources with different protein content on colony performance, wintering ability and in-vitro longevity of colonies that weakened after feeding with pine honey in autumn or that needed to enter the winter period were investigated. The experiment was carried out in 48 colonies divided into six groups as follows: control, syrup, mixed pollen, Cistus creticus pollen (Pink rock-rose), Papaver somniferum pollen (Opium poppy), and commercial bee cake group. The effect of nutritional differences on survival was found to be statistically significant in vitro and this supports the colony results in the natural environment. As a result, P. somniferum pollen is a good preference to be used in feeding colonies in beekeeping, due to its rich nutritional content.Honey bees need pollen and nectar sources to survive in nature. Particularly, having young bees in colonies is vital before wintering, and proper feeding is necessary to achieve this. In the present study, the effect of feeding with pollen sources of different protein content on colony performance, wintering ability and in-vitro longevity of colonies that weakened after feeding with pine honey in autumn, or that needed to enter the winter period, was investigated. The experiment was carried out in 48 colonies divided into six groups as follows: control, syrup, mixed pollen, Cistus creticus pollen (Pink rock-rose), Papaver somniferum pollen (Opium poppy), and commercial bee cake groups. In particular, the P. somniferum pollen group was different (p < 0.01) from the other experiment groups with the number of bee frames (3.44), the area with brood (1184.14 cm2) and the wintering ability of 92.19%. The effect of nutritional differences on survival was found to be statistically significant in vitro and this supports the colony results in the natural environment (p < 0.001). The P. somniferum group has the longest longevity with 23 days. Pollen preferences of honey bees were P. somniferum, C. creticus, and mixed pollen, respectively.

Composition of Essential Oils in Needles and Barks of Turkish Red Pine (Pinus brutia Ten.) Infested by Marchalina hellenica Genn.

The scale insect Marchalina hellenica Genn. plays a key role in pine honey production and is hosted mainly by Turkish red pine (Pinus brutia Ten.). The needles of Turkish red pine are rich in essential oils. Moreover, essential oils can affect the host selection of insects. The essential oils in the needles and barks of Marchalina hellenica-infested and non-infested Turkish red pine were obtained via water distillation and their composition was determined by GC-MS analyses. The composition of the essential oils was found to differ in the needles of non-infested Turkish red pine and in those of pine infested by Marchalina hellenica. The Mann-Whitney U test results showed that the β-caryophyllene level was higher in the needles of Turkish red pine infested by Marchalina hellenica, whereas the junipen level was higher in the essential oil of non-infested Turkish red pine bark. Pimaral and α-guaiene were detected only in the needles of infested trees, but were absent in the needles of non-infested trees. These components may be markers that can act on their own or as part of the whole in the host tree selection of Marchalina hellenica. In addition, the cis-verbenone component found in Turkish red pine bark might play a role in attracting Marchalina hellenica. This phenomenon should be further investigated through additional studies.