rss_2.0Journal of Apicultural Science FeedSciendo RSS Feed for Journal of Apicultural Science of Apicultural Science Feed of Propolis Commercially Available on Podkarpacki Beekeeping Market<abstract> <title style='display:none'>Abstract</title> <p>Eight samples of propolis commercially available on south-eastern Poland’s beekeeping market were compared in terms of quality, chemical composition and biological activity, including antioxidant and antimicrobial properties. The organoleptic characteristics and physicochemical requirements indicated that only 50% of tested propolis samples was classified to class II regarding obligatory limits, while the others were out-class. A big variation in mineral composition was determined by the ICP-OES method but all samples were free of heavy metals (Pb, Cd and Ni). Although mesophilic bacteria, yeasts and moulds occurred in tested propolis (in general lower than 10<sup>2</sup> CFU/g), <italic>E. coli</italic> and spore-forming sulphite-reducing bacteria as well as <italic>Salmonella</italic> and <italic>Shigella</italic> bacteria were not detected. High antioxidant activity and total phenolic content for all samples was confirmed, which was correlated with antibacterial activity tested against <italic>E. coli</italic> ATCC 11775 strain. Additionally, the propolis sample with the strongest antibacterial activity (MIC 0.33 mg/mL) inhibited biofilm formation of <italic>S. aureus</italic> and <italic>S. epidermidis</italic> ATCC 35984 (MBIC 0.66 and 5.25 mg/mL, respectively). However, great variation in terms of antioxidant activity and HPLC polyphenolic profile were observed, and sakuranetin and pinobanksin were identified as the most effective antioxidant components. Moreover, the usefulness of ATR-FTIR spectra analysis was confirmed as a quick method for initial testing of propolis quality.</p> </abstract>ARTICLEtrue of Willow Rust Spores ( Spp.) Collected by Honey Bees<abstract> <title style='display:none'>Abstract</title> <p>Honey bee workers (<italic>Apis mellifera</italic> L.) collect nectar, honeydew and pollen from plants in orderto provide the colony with, among others, carbohydrates and protein. Whenever these sources are unavailable in the environment, bees turn to alternatives. An example of this are fungi spores collected accidentally or on purpose. This last phenomenon is the aim of this study, in which we have shown that worker bees can collect willow rust (<italic>Melampsora</italic> spp.) spores. We observed as the bees obtained spores and placed them in pollen baskets. The presence of spores was demonstrated with the use of a scanning electron microscope (SEM). This observation indicates that the honey bee may use alternative sources as a potential supplement. There are few studies in this field and it requires a deeper analysis.</p> </abstract>ARTICLEtrue Affiliation of in Selected Locations from Poland and Ukraine<abstract> <title style='display:none'>Abstract</title> <p>The virulence patterns observed in viruses transmitted by <italic>Varroa destructor</italic> implied a potential association of these honey bee pests in Poland with the Korean haplotype. Nevertheless, it is noteworthy that this hypothesis lacked direct verification. Therefore, the aim of the study was to determine the haplotype affiliation of <italic>V. destructor</italic> mites infesting bees in Poland and Ukraine. Adult females were collected from drone brood from honey bee colonies maintained in the apiary at the University of Life Sciences in Lublin, Poland, and the apiary in Synevyr National Park in Ukraine. The haplotype affiliation was determined through the sequencing of the cytochrome oxidase subunit I gene (CO I; 929 bp) fragment. Based on the phylogenetic comparison with 84% bootstrap support, the <italic>V. destructor</italic> females from both locations were confirmed to represent three haplotypes: Korean 1 (AmK1-1), Korean 2 (AmK1-2), and Chinese 4 (AmK1-4). They are the most virulent types of this parasite worldwide and in Poland. Concurrently, the present study confirms that the Synevyr National Park in Ukraine is isolated from alien bee subspecies, but is not free from alien parasites.</p> </abstract>ARTICLEtrue and Shape Differences in Fore Wings of Honey Bee () Queens, Workers and Drones<abstract> <title style='display:none'>Abstract</title> <p>The three honey bee phenotypes - queens, workers, and drones differ in the size and shape of body parts. We used a large dataset to describe how the three phenotypes differed with regard to fore wings and applied the methodology of geometric morphometrics to measure the wings using the coordinates of nineteen landmarks. On average, queens had wings larger than workers, but the two castes had a broad overlap. Drones, on the other hand, had distinctly larger wings. Wing shape differed markedly among queens, workers and drones and can be used to reliably differentiate them from one other. Surprisingly, the fore wing shape of workers was more similar to that of drones than to that of queens. Small queens were not more similar to workers than large queens, and large workers were not more similar to queens than small workers. Because wing size, unlike body mass, does not change over the life of the queen, it can be used to evaluate the quality of a queen throughout its life and not only during a short period after emergence. We provide a large number of wing images of queens and drones, which can be used as a reference in future studies.</p> </abstract>ARTICLEtrue of in Africanized and European Honey Bee, , Lineages from the United States<abstract> <title style='display:none'>Abstract</title> <p>Honey bee populations in the United States have been under stress for the past several decades. Several internal parasites may contribute to this, including the trypanosome <italic>Lotmaria passim</italic>. It is unknown how widespread the honey bee parasite, <italic>L. passim</italic>, is in Africanized honey bee (AHB) populations in the United States. A total of 321 feral honey bee colony samples which had been previously recorded to be of Africanized origin using molecular diagnostics from California (n=3), New Mexico (n=46), Oklahoma (n=57), Texas (n=106), and Utah (n=109) were used in this study. Of these samples, a total of 15 (4.7%) from three States were positive for <italic>L. passim</italic>. Utah AHB samples had the highest infection rate (11.0%), followed by Texas (1.9%) and Oklahoma (1.8%). Compared with previous studies on the occurrence of <italic>L. passim</italic> from European honey bees from the same sampled States, infection rates of Africanized honey bees for <italic>L. passim</italic> were significantly higher in the State of Utah, but not for the other sampled States. This study provides evidence that feral honey bee populations do not necessarily have lower levels of honey bee parasites than managed honey bee colonies.</p> </abstract>ARTICLEtrue Pollination and Pollinators in Farming of Insecticides with Different Modes of Action to Larvae (Hymenoptera, Apidae)<abstract><title style='display:none'>Abstract</title> <p>Pesticides are one of the elements that lead to the decline in honeybee populations. Honey bee while foraging collect nectar and pollen from various crops, can be exposed to pesticide residues, which may be transported into their colonies and consumed by the larvae. The objective of this study was to evaluate acute toxicity of fipronil and lambda-cyhalothrin, two commonly detected pesticides within beehives, on the larvae of <italic>Apis mellifera</italic>. The testes chemicals were given to three-day old larvae. Treatments were administered using a micropipette at the bottom of each chosen comb cell. Each treatment included sixty larvae per dosage and the experiment was repeated three times. Larvae were given only acetone in the control treatment. Fipronil and lambda-cyhalothrin were toxic to the larvae of <italic>A. mellifera</italic> with LD<sub>50</sub> values of 0.163 and 0.83 μg/larvae, respectively after seven days of exposure. The maximum mortality of 81.35% was recorded with a dose of 0.38 μg/larvae in Fipronil, whereas in lambda-cyhalothrin, it was 94.23% with a dose of 1.70 μg/larvae.</p> </abstract>ARTICLEtrue of Pollination by the (syn. ) Bee on Fruit Set, Seed Set and Yield in Three Apple Cultivars<abstract><title style='display:none'>Abstract</title> <p>Our study tested the pollination effectiveness of the megachilid bee <italic>Osmia bicornis</italic> L. (syn. <italic>Osmia rufa</italic>) on three apple cultivars (cv.): Lobo, Piros and Champion. The following pollination treatments were compared for each cultivar: insect pollination - caged trees with <italic>Osmia bicornis</italic> (syn. <italic>Osmia rufa</italic>) bees; open pollination - uncaged trees; self-pollination - caged trees without bees. Pollination success was measured based on the percentage of fruit set, the fruit yield per tree (measured as number of fruits and total fruit weight) and the number of seeds per fruit. Generally, there was a higher percentage of initial fruit-set and fruits at harvest on open-pollinated trees than on caged trees without bees, with an intermediate value recorded for caged trees pollinated by <italic>O. bicornis</italic> (syn. <italic>O. rufa</italic>). However, the fruit set on trees pollinated by <italic>O. bicornis</italic> (syn. <italic>O. rufa</italic>) was high and ranged from 49 to 69%, depending on the cultivar. The cv. Lobo trees pollinated by <italic>O. bicornis</italic> (syn. <italic>O. rufa</italic>) and open-pollinated trees produced higher fruit yields than without bees. Furthermore, the number of seeds per fruit was higher. Fruit yield and the number of seeds per fruit were higher for trees pollinated by <italic>O. bicornis</italic> (syn. <italic>O. rufa</italic>) than for caged trees without insects but lower than for open pollination trees of cv. Piros and cv. Lobo.</p> </abstract>ARTICLEtrue Count Dynamics in Rapeseed Stamens in Early Spring<abstract><title style='display:none'>Abstract</title> <p>Pollen contains vegetative and generative cells that influence the seed set. Accurate pollen assessments help understand the breeding biology of oilseed rape. In order to evaluate the number of pollen grains in a large number of samples, an effective method should be designed. A new method was established to evaluate the pollen counts of five oilseed rape cultivars. This method mainly relies on 5% sucrose + 0.1% agar solution to suspend the pollen and uses a standard curve corresponding to the absorbance and the number of pollen grains to estimate the number of pollen grains. The linear fit formula of this standard curve was precision, and the R<sup>2</sup> value between the pollen count and absorbance reached 0.998. Furthermore, the variation in the pollen counts at three flowering stages and on two kinds of stamens was assessed. The pollen count per flower varied significantly among oilseed rape cultivars and flowering stages (GLM: p&lt;0.001). Moreover, the number of pollen grains on long-stamen anthers was significantly higher than that on short-stamen anthers (GLM: p&lt;0.001). In conclusion, we established a rapid, accurate method for quantifying pollen grains based on absorbance determined in a liquid suspension by light spectroscopy, which is a feasible method. In addition, the number of pollen grains under different physiological conditions also provides basic data for oilseed rape breeding.</p> </abstract>ARTICLEtrue Defence Strategies of the Stingless Bee, Smith. Against Nest Intruders in a Newly Divided Colony<abstract><title style='display:none'>Abstract</title> <p>A study was conducted to document nest intruders in newly divided colonies of stingless bees. The nature of the damage, the sequence of activities after colony division, and task allocation in the colony were studied. Three insect invaders - solitary resin bees (<italic>Megachile disjuncta</italic>, <italic>M. cephalotes</italic>), ants (<italic>Oecophylla smaragdina</italic> and <italic>Camponotus</italic> sp.) and spiders belonging to the family Salticidae and Lycosidae invaded the colonies during the earlier stages of division. The mean number of resin bees and ant invasions was significantly higher during the first week after division. Spiders invaded the hives and formed webs during the second week after division, resin and honey resources are robbed by resin bees and ants. Nest entrance closure was observed at 4.37±0.74 days after colony division guard bee activity from 6.13±1.24 days and foraging activity from 10.63±1.06 days after division. Observations on the task allocation framework after colony division indicated that 69% of bees were involved in hive entrance repair, 18% in colony architecture development, and 13% in tending to newly emerged bees from brood cells. The bees built a complex nest entrance, guarded activity and demonstrated nest closure behaviour in order to protect the nest against intruders. The results of the study are of prime importance for beekeepers to learn how to watch for hive intruders during colony division and to understand the adaptive defence mechanism to protect colonies.</p> </abstract>ARTICLEtrue of Diversity and Composition of Gut Microbiome in Overwintering Period<abstract><title style='display:none'>Abstract</title> <p>Cold temperatures limit the survival and reproduction of honeybees. Therefore, successful overwintering is crucial for the survival of honeybee colonies and the pollination of flowering plants in the following spring. This study analyzed the gut microbiota of <italic>Apis cerana</italic> from Changbai Mountain during the overwintering period through 16S rDNA high-throughput sequencing. The analysis of microbial community composition showed that <italic>Gilliamella</italic>, <italic>Lactobacillus</italic>, <italic>Bifidobacterium</italic> and <italic>Snodgrassella</italic> were the core genera in the honeybee gut during the overwintering period. Moreover, alpha diversity analysis showed that the community diversity of the gut microbiota peaked in December. Beta diversity and LEfSe analysis showed that community composition was similar in December, January and February and that such beneficial bacteria as <italic>Snodgrassella</italic>, <italic>Acetobacteraceae</italic>, and <italic>Rhizobiales</italic> were differentially abundant during this period. The results of functional prediction analysis indicate that amino acid transport and metabolism, carbohydrate transport and metabolism, and the TCA cycle play an important role in overwintering. These results suggest that the dynamic changes in the gut microbiota of <italic>A. cerana</italic> during winter and mechanisms tolerate cold stress.</p> </abstract>ARTICLEtrue of Electromagnetic Field with Frequency of 50 Hz in form of Doses on Selected Biochemical Markers of Honey Bee<abstract> <title style='display:none'>Abstract</title> <p>The honey bee has a significant environmental and economic impact. While collecting food or water, bees are exposed to negative physical and chemical factors that lead to physiological and behavioral changes and, consequently, even death. Along with the development of technology and communication, electromagnetic fields produced by artificial emitters have begun to have an impact on the environment. The aim of the study was to check whether the electromagnetic field also impacts antioxidant enzymes functioning in the honey bee's organism. The study was conducted under laboratory conditions, and one-day and seven-day-old honey bee workers were used in the experiment. Honey bee workers were exposed to an electromagnetic field with a frequency of 50Hz and variable intensity in the range of 1–10 kV/m. Immediately after the end of the exposure, hemolymph was collected from the bees for biochemical analysis. The results of the research did not show clearly whether changes in the activity of biochemical markers were affected by the time spent in the electromagnetic field or its intensity but did show that there was a difference in physiology between one-day-old and seven-day-old bees.</p> </abstract>ARTICLEtrue and Molecular Characterization of Deformed Wing, Acute Bee Paralysis and Black Queen Cell Viruses Infecting Honey Bees and Mites<abstract> <title style='display:none'>Abstract</title> <p>Surveys were conducted in Malatya and Elazığ provinces of Eastern Türkiye between 2018–2019 to detect the presence and distribution of <italic>Deformed wing virus</italic> (DWV), <italic>Acute bee paralysis virus</italic> (ABPV) and <italic>Black queen cell virus</italic> (BQCV) infecting honey bees and <italic>Varroa</italic> mites. Thirty <italic>Varroa</italic> mites feeding on honey bees and 147 honey bees were collected from twenty-five apiaries of Malatya and fifteen apiaries of Elazığ. The collected samples were subjected to reverse transcriptase-polymerase chain reaction. All tested viruses were found to be present in the samples singly or as mixed infections. The prevalence of the viruses were 23.81%, 12.93%, and 10.20% for DWV, ABPV and BQCV, respectively. Five out of thirty <italic>Varroa</italic> mites tested for the viruses were found to be infected only with DWV. Randomly chosen sequences of each detected virus’s partial polyprotein gene region were registered in GenBank under the accession numbers OP805878, OP805879, OP805880, OP805887, OP805888, OP805889, OP805890, OP805891 (DWV), OP805881, OP805882, OP805883, OP805884 (ABPV) and OP805885, OP805886 (BQCV). The phylogenetic tree of the viral isolates were compared with the world isolates. DWV isolates were found to be closely related to the UK, Lebanon, Türkiye, France, Germany and Israel isolates, whereas ABPV isolates were related to Türkiye, South Africa, Slovenia, Serbia, France, Hungary, Syria and USA isolates and BQCV isolates were related to China, South Korea, Czech Republic, France, Hungary, Poland, South Africa, Australia and Sweden isolates. The presence of DWV, ABPV and BQCV infections in Malatya and Elazığ provinces of Türkiye is revealed for the first time in this study.</p> </abstract>ARTICLEtrue Study on Physicochemical, Antioxidant and Antimicrobial Properties of Propolis Collected from Different Regions of Bulgaria<abstract> <title style='display:none'>Abstract</title> <p>Propolis (bee glue), a valuable biological mixture produced by European honey bees (<italic>Apis mellifera</italic> L.), is widely used as a remedy in traditional and alternative medicine, as an effective food biopreservative and as a nutritional value enhancer. The present study aimed to investigate eighty propolis samples collected from all twenty-eight districts of Bulgaria in the period of 2020–2022 in order to determine their physicochemical properties as well as their antioxidant and antimicrobial potential. The raw propolis samples exhibited brown or green colour depending on the climatic zone from which they are collected and pH values from 4.82 to 5.87, while the moisture content ranged between 0.98% and 2.97%. The total phenolics content (TPC) and total flavonoids content (TFC) varied from 63.14 mg GAE/g to 737.27 mg GAE/g, and from 29.22 mg QE/g to 234.17 mg QE/g, respectively. The propolis samples demonstrated significant antioxidant potential determined by DPPH and FRAP methods, from 18.56 mM TE/g to 1598.66 mM TE/g and from 82.28 mM TE/g to 1208.81 mM TE/g, respectively, whose values showed a positive correlation (<italic>r</italic><sup>2</sup>) with the TPC and TFC. The results from the antimicrobial screening revealed that the methanolic propolis extracts in concentration of 20 mg/ml exhibited the highest inhibitory effect on the Gram-positive bacteria such as <italic>Micrococcus luteus</italic> 2YC-YT, <italic>Bacillus subtilis</italic> ATCC 6633, <italic>Staphylococcus aureus</italic> ATCC 25923, <italic>Listeria monocytogenes</italic> NBIMCC 8632 and <italic>Listeria innocua</italic> ATCC 33090.</p> </abstract>ARTICLEtrue between Flower Opening Time, Environmental Conditions, Corolla Opening Size and Nectar Production in Five Winter Oilseed Rape ( L.) Cultivars in China<abstract> <title style='display:none'>Abstract</title> <p>Oilseed rape is an economically important crop and provides a good nectar resource. In this study, the nectar secretion characteristics of five winter oilseed rape (<italic>Brassica napus</italic> L.) cultivars in China were examined throughout the daytime. The correlations of corolla opening size, air temperature and relative humidity with nectar production were also determined. Different daily patterns of nectar secretion were detected in flowers opening at different times of the day, and similar trends were observed among cultivars. The nectar volume and nectar sugar concentration in flowers that opened before 9:00 peaked on the first day. The nectar volume of flowers that opened at 11:00 peaked on the second day, and the nectar sugar concentration peaked on the first day. The total nectar yield of flowers opening before 9:00 (4.422–5.265 μl) was lower than that of flowers opening at 11:00 (7.982–10.646 μl). The average nectar sugar concentration of flowers opening before 9:00 (18.4–23.3%) was higher than that of flowers opening at 11:00 (15.3–17.5%). The nectar volume was positively correlated with air temperature and relative humidity, while the nectar sugar concentration was positively correlated with air temperature and negatively correlated with relative humidity. The nectar volume and nectar sugar concentration were positively correlated with corolla opening size. The glucose/fructose ratio ranged from 0.89 to 1.44. These results will contribute to a better understanding of the nectar secretion patterns of <italic>B. napus</italic> L. and will support further studies assessing the melliferous potential of <italic>B. napus</italic> L. and the temporal dynamics of plant-pollinator interactions.</p> </abstract>ARTICLEtrue Status of Honeybee Colonies Differing in Genetic Intra-Colonial Diversity<abstract> <title style='display:none'>Abstract</title> <p>Two different levels of diversity within a colony were compared for the prevalence of pathogens and diseases. Lower genetic diversity was obtained in the colonies in which the queens were inseminated with semen collected from drones originating from a single colony, while greater was obtained in the colonies with queens inseminated with semen from drones of thirty different colonies. Bees were tested for <italic>Varroa destructor</italic> infestation, microsporidia <italic>Vairimorpha</italic> spp. infection, acute bee paralysis virus (ABPV) and deformed wing virus (DWV). Colonies with a greater genetic diversity of workers in colonies were more infested with <italic>Varroa</italic> mites than genetically uniform colonies. <italic>Varroa</italic> infestation was not found to be associated directly with the weakening of bee colonies after winter. The two experimental groups had a similar number of colonies infected with <italic>Vairimorpha</italic>, and viruses. Intensity of <italic>Varroa</italic> infestation and <italic>Vairimorpha</italic> infection did not significantly affect the overwintering of bee colonies. Colonies in which DWV was detected significantly weakened during overwintering.</p> </abstract>ARTICLEtrue of Mercury Level in Local Polish and Imported Honeys with Use of Direct Mercury Analyzer<abstract> <title style='display:none'>Abstract</title> <p>Mercury is a toxic metal which causes several serious health effects including kidney damage, anxiety depression and peripheral neuropathy, and because of its high volatility assessment in solid samples is problematic. In this study, forty-five samples of local Polish and imported honeys were analyzed by Direct Mercury Analyzer (DMA) for the determination of mercury content. The honey contamination was assessed regarding honey variety and country of origin. Hg level in all tested samples was found to not exceed the 0.81 μg/kg which was below 10% of the applicable law UE limit. Moreover, no statistically significant differences were found in the variety or geographical origin. The study concluded that the Direct Mercury Analyzer (DMA) instrument proved to be an excellent tool for screening mercury in honeys because of its simplicity, rapidity, low detection limit, accuracy and precision and no sample preparation.</p> </abstract>ARTICLEtrue of Entomophily and Artificial Pollination in Enhancing Quality and Yield of Seed Onion ( L.) in Indian Himalayas<abstract> <title style='display:none'>Abstract</title> <p>Onion (<italic>Allium cepa</italic> L.) is a highly cross pollinated crop that needs insect pollination (entomophily) for optimum seed set. In the present study, more than 120 insect species were noted to visit onion flowers, out of which fifty-nine species were collected through in-situ sampling technique and identified up to species level. The Indian bee (<italic>Apis cerana indica</italic>) was the most abundant insect pollinator visiting onion flowers in the study location. The pollination behaviour and foraging activity of <italic>A. c. indica</italic> were assessed, and the pollen foragers were observed to be swift flyers visiting a significantly higher number of flowers per minute during the peak flowering period and spending less time per flower (3 seconds) to collect nectar from deep seated nectaries of the flowers. The seed yield enhancement assessed through entomophily and artificial pollination methods showed that the open pollinated flowers recorded the highest yield statistically, followed by <italic>A. c. indica</italic> and <italic>A. mellifera</italic> pollinated flowers. However in artificial pollination treatments, sponge puff pollinated flowers recorded significantly high seed yield per hectare and percentage seed set per umbel, followed by camel brush, hand gloves (cloth) and hand gloves (rubber). In conclusion, all the cross pollination treatments were statistically significant on the closed pollination treatment concerning to the entire yield parameters calculated.</p> </abstract>ARTICLEtrue of Oxytetracycline on Colonies: Preliminary Results on Residues and Antibiotic Resistance<abstract> <title style='display:none'>Abstract</title> <p>We tested two summer protocols for oxytetracycline (OTC) administration on beehives to evaluate: toxicity for the bees, residues in honey and presence of genes encoding for OTC antibiotic resistance in bacteria isolated from the gut of adult bees. In July 2020, eighteen healthy colonies were divided into three groups. Two OTC treatment protocols - long and short, were devised, and one group was left untreated. We did not detect any toxicity on adult bees or brood but found OTC residues in the honey of the nest up to seven months after both treatments (210.3±221.2 µg/kg after long protocol; 216.2±238.4 µg/kg after short protocol), even in the untreated group (up to the mean 75.8±173.2 µg/kg). Antibiotic resistance genes were found, even in the untreated group, possibly due to the long exposure to antibiotics. More in-depth studies should be performed to verify how the long-term persistence of antibiotics impacted the beehive during the inactive season.</p> </abstract>ARTICLEtrue Water Vapor Sorption Properties of Honey<abstract> <title style='display:none'>Abstract</title> <p>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 (a<sub>w</sub>)&gt;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&gt;0.05). The honeys exhibited a sorption isotherm (SI) in the shape of a “J”. Their SIs exhibited desorption and adsorption at a<sub>w</sub>&lt;0.7 and a<sub>w</sub>&gt;0.7, respectively and the desorption part was almost linear. A<sub>w</sub>=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&gt;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.</p> </abstract>ARTICLEtrue