Gamma irradiation and ozone application as preservation methods for longer-term storage of bee pollen.

Bee pollen is a healthy product with a good nutritional profile and therapeutic properties. Its high moisture content, however, promotes the growth of bacteria, molds, and yeast during storage commonly result in product degradation. Therefore, the aim of this study is to assess the effectiveness of gamma irradiation (GI) and ozone (OZ) as bee pollen preservation methods for longer storage time, as well as whether they are influenced by pollen species. To do that, GI at a dosage of 2.5, 5.0, and 7.5 kGy was applied at a rate of 0.68 kGy/h and OZ application at a concentration of 0.01, 0.02, and 0.03 g/m3 was applied for one time for 6 h, to Egyptian clover and maize bee pollen, then stored at ambient temperature for 6 months. We then determined the total phenolic content (TPC) and antioxidant activity of treated and non-treated pollen samples at 0, 3, and 6 months of storage. Total bacteria, mold, and yeast count were also evaluated at 0, 2, 4, and 6 months. Statistical analyses revealed that, TPC, antioxidant, and microbial load of both clover and maize pollen samples were significantly (p < 0.05) affected by both treatment and storage time and their interaction. Both methods were extremely effective at preserving the antioxidant properties of pollen samples after 6 months of storage at room temperature. Furthermore, the highest concentrations of both GI and OZ applications completely protected pollen samples from mold and yeast while decreasing bacterial contamination. GI at the highest dose (7.5 KGy) was found to be more effective than other GI doses and OZ application in preserving biologically active compounds and lowering the microbial count of pollen samples for 6 months. As a result, we advise beekeepers to use GI at this dose for longer-term storage.

Combining Ability and Inheritance Nature of Agronomic Traits and Resistance to Pink Stem (Sesamia cretica) and Purple-Lined (Chilo agamemnon) Borers in Maize.

The pink stem borer (PSB), Sesamia cretica (Lepidoptera: Noctuidae) purple-lined borer (PLB), Chilo agamemnon (Lepidoptera: Crambidae) and European corn borer Ostrinia nubilalis, (Lepidoptera: Crambidae) are considered the most devastating insect pests of maize production in the Mediterranean region. The frequent use of chemical insecticides has resulted in the evolution of resistance to various insect pests as well as the pernicious impact on natural enemies and environmental hazardousness. Therefore, developing resistant and high-yielding hybrids is the best economic and environmental approach to cope with these destructive insects. Accordingly, the objective of the study was to estimate the combining ability of maize inbred lines (ILs), identify promising hybrids, determine gene action controlling agronomic traits and resistance to PSB and PLB, and investigate inter-relationships among evaluated traits. A half-diallel mating design was employed to cross seven diverse maize inbreds to generate 21 F1 hybrids. The developed F1 hybrids, alongside high-yielding commercial check hybrid (SC-132), were assessed in field trials for two years under natural infestation. Substantial variations were obtained among the evaluated hybrids for all recorded characteristics. The non-additive gene action was major for grain yield and its contributing traits, while the additive gene action was more important in controlling the inheritance of PSB and PLB resistance. The inbred line IL1 was identified to be a good combiner for earliness and developing short-stature genotypes. Additionally, IL6 and IL7 were recognized as excellent combiners to enhance resistance to PSB, PLB and grain yield. The hybrid combinations IL1×IL6, IL3×IL6, and IL3×IL7 were determined to be excellent specific combiners for resistance to PSB, PLB and grain yield. Strong positive associations were identified among grain yield, its related traits, and resistance to PSB and PLB. This implies their importance as useful traits for indirect selection for improving grain yield. Otherwise, the resistance against PSB and PLB was negatively associated with the silking date, indicating that earliness would be favorable for escaping from the borer's attack. It could be concluded that the inheritance of PSB and PLB resistance can be governed by the additive gene effects, and the IL1×IL6, IL3×IL6, and IL3×IL7 hybrid combinations can be recommended as excellent combiners for resistance to PSB and PLB and good yield.

Influence of Effective Microorganisms on Some Biological and Biochemical Aspects of Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae).

The cotton leafworm, Spodoptera littoralis (Bosid.), is a major pest in African and Asian nations that attacks a wide variety of host plants. This study was conducted to assess the effectiveness of effective microorganisms (EMs) on the biological and physiological features of S. littoralis larvae. Five concentrations (100, 200, 300, 400, and 500 ppm) of EMs were tested. Antifeedant activity, food consumption index, the efficiency of converting digested food, the efficiency of converting ingested food, relative growth rate, and approximate body tissue of the fourth larval instar of S. littoralis were determined. Moreover, carbohydrate enzyme activities (amylase, trehalose, and invertase), total protein, and total lipids of S. littoralis larvae were measured to elucidate the mode of action of the tested agent in the S. littoralis's larval stage. The EMs at 500 ppm had a substantial impact on antifeedant activity, nutritional indices, egg deposit reduction, and hatchability in S. littoralis during the five days. All concentrations interrupted S. littoralis's life cycle and developmental phases. Furthermore, all concentrations were quite useful in lengthening the developmental stages of S. littoralis. In addition, Ems affected the biochemical activities of larvae, leading to disturbances in carbohydrate, lipid, and protein levels. From this study, EMs can be used as a bioinsecticide alternative to traditional insecticides against S. littoralis and may be compatible with integrated pest management approaches.

Gamma Radiation: An Eco-Friendly Control Method for the Rice Weevil, Sitophilus oryzae (L.) (Coleoptera: Curculionidae).

The rice weevil, Sitophilus oryzae (L.), is the most destructive insect pest of stored cereals worldwide. The current study was conducted to determine the lethal, reproductive, and histological effects of gamma irradiation on S. oryzae adults. In addition, the impact on germination, chlorophyll, and proline content in wheat seedlings from treated grains was determined. Wheat grains were infested with rice weevil adults and then irradiated by gamma rays. Gamma radiation was applied at a dosage of 0.10, 0.25, 0.50, and 1.00 kGy. Mortality percentage and LD50 were recorded after 48, 72, 96, and 120 h of treatment. The dosage of 1.00 kGy caused 100% mortality after 96 h of irradiation. The required dosage of gamma radiation to kill 50% (LD50) of adults after 48 h was 1.51 kGy. All tested doses caused complete sterility to 24 h old adults. A histological alteration was noticed at a dosage of 1.00 kGy, which showed cytoplasmic vacuolization, tissues exhibiting signs of putrefaction, and necrosis of cells; furthermore, gamma irradiation affected chlorophyll a and b. The highest amounts were detected in wheat seedlings from grains irradiated at 0.10 kGy. There was a significant increase in plant proline content at the higher doses (0.50 and 1.00 kGy) compared with seedlings from nonirradiated grains. It could be concluded that gamma radiation can be used as an eco-friendly trend to control stored-product pests without any residual effects.

The impact of cold storage durations on Trichogramma evanescens (Westwood) (Hymenoptera: Trichogrammatidae) during their pupal stage.

This experimental study was done at the Biological Control Laboratory, Sakha Agricultural Research Station, Sakha, Kafrelsheikh, Egypt. We aimed to estimate the impact of different cold (10 °C) storage durations [0 (non-cold-stored parasitized eggs), 3, 6, 9, 12, 15, 18, 21, 24, 27, and 30 days], on Trichogramma evanescens (Westwood) (Hymenoptera: Trichogrammatidae) during the pupal stage using the eggs of Sitotroga cerealella after exposing to T. evanescens. The emergence percentage of non-cold-stored eggs of S. cerealella was higher than all cold-stored durations. Also, the female's percentages of T. evanescens in the cold storage durations were lower than the non-cold storage one, and they were influenced by extended cold storage durations. There were non-significant differences in the female's longevity of T. evanescens obtained from 0, 3, and 6 days cold-stored parasitized eggs of S. cerealella at 10 °C, but it began to decrease from those produced after 9 days of cold-stored eggs. In addition, the emergence percentage in F1 progeny of T. evanescens was greater than 50% until 21 days of cold storage. It could be concluded that cold storage reduced the % emergence, % females, female's longevity, and emergence percentage in F1 progeny of T. evanescens. For a successful biological control program, the decrease of T. evanescens performance after cold storage durations should be considered in mass production, and the release percentage should be increased by the equivalent of a lack of % emergence. Also, the economic importance of using cold storage periods in commercial mass rearing should be assessed in the biological control program.

Effect of comb age on cell measurements and worker body size.

The honey bees (Apis mellifera L.) mainly use beeswax (comb) for brood rearing and food storage. Changes in the color and cell dimensions occur due to repeated food storage and brood rearing in the comb. The study aimed to determine the changes in comb cell measurements and worker body sizes in relation to comb age. For this purpose, the cell measurements of combs at age zero (wax foundation), 1, 2, 3, 4, 5, and 6 years and the body size of workers reared in them were estimated. The weight of the comb, the height of the cell base, and the weight of accumulated substances in the cell significantly increased with time. Comb age had negative effects on the cell diameter, cell depth, cell volume, cell honey or pollen capacity, and newly emerged worker body weight. Significant negative correlations were observed between the accumulated substances in a cell and the cell diameter, cell depth, and cell size, while significant positive correlations were observed among the cell volume, cell diameter, cell depth, cell honey capacity, cell pollen capacity, and worker body weight. It can be concluded that the dimensions of the comb cells and worker body size changed with the age of the comb. The obtained results recommend beekeepers to replace combs aged more than 3 years with a new comb to allow large workers to gather more nectar and pollen, rear a larger brood, and store more honey.

Harvest Season Significantly Influences the Fatty Acid Composition of Bee Pollen.

Seasonal variations in the fatty acid (FA) compositions of pollen loads collected from the Al-Ahsa Oasis in eastern Saudi Arabia throughout one year were determined to identify the optimal season for harvesting bee pollen rich in essential fatty acids (EFAs) and unsaturated fatty acids (UFAs). The highest values (%) of lipids, linolenic acid (C18:3), stearic acid (C18:0), linoleic acid (C18:2), arachidic acid (C20:0), the sum of the C18:0, C18:1, C18:2, and C18:3 concentrations, and EFAs were obtained from bee pollen harvested during autumn. The maximum values (%) of oleic acid (C18:1), palmitic acid (C16:0), UFAs, and the UFA/saturated fatty acid (SFA) ratio were found in bee pollen harvested during summer. The highest concentrations (%) of behenic acid (C22:0), lignoceric acid (C24:0), and SFAs were found in bee pollen harvested during winter. Bee pollen harvested during spring ranked second in its oleic, palmitic, linolenic, stearic, arachidic, behenic, and lignoceric acid concentrations and for EFAs, UFAs, and the UFA/SFA ratio. The lowest SFA concentration was found in bee pollen harvested during summer. Oleic, palmitic, and linolenic acids were the most predominant FAs found in bee pollen. It was concluded that the FA composition of bee pollen varied among the harvest seasons due to the influence of the dominant botanical origins. We recommend harvesting pollen loads during spring and summer to feed honeybee colonies during periods of scarcity and for use as a healthy, nutritious food for humans.

The impact of caging the queens during the flow season on some biological activities of honeybee colonies.

This study was achieved in a private apiary located in a banana farm in Sa El Hagar, Basioun, Gharbia, Egypt from August 15, 2019 to May 25, 2020, including the banana (Musa sp., Musaceae) flow season (August and September) and extend to Egyptian clover (Trifolium alexandrinum L., Fabaceae) flow season (May). The study aimed to evaluate the effect of confining the queen during the banana flow season on the brood rearing, honey yield, and activation of worker's ovaries. Also, we determined the negative impact of caging the queen during the banana flow season on the activity of the colony in brood rearing, storing pollen, and honey yield after releasing the queen on 5 October, extending to the next flow season in May. The obtained results showed that the honeybee colonies with the caged queen produced significantly more honey yield and less brood production than the free queen ones during the banana flow season. Also, the caging of the queen did not affect the colony strength after releasing the queen despite the partial development of the ovaries of some workers, but they did not lay eggs. In addition, releasing the queens suppressed the ovaries of the laying workers. It can be concluded that caging the queen during the banana flow season helps the colonies to produce more honey yield without effect on the colony strength after releasing the queen despite the ovaries development of few workers without egg-laying.

Morphometric study of Yemeni (Apis mellifera jemenitica) and Carniolan (A. m. carnica) honeybee workers in Saudi Arabia.

The Yemeni honeybee (Apis mellifera jemenitica Ruttner) is the native race in Saudi Arabia. The Carniolan honeybee (A. m. carnica Pollmann) and its hybrid with the Egyptian honeybee (A. m. lamarkii Cockerell) have been imported and frequently reared in Saudi Arabia. Temperature often exceed 40 °C during the summer season in most regions of Saudi Arabia. Honeybees decrease foraging activity in this period during mid-day, which affect colony productivity. The Yemeni bee race appears well adapted to these unique climatic conditions. We compared body weight and morphometric parameters of both subspecies' worker bees reared at the apiary of Training and Research Station, King Faisal University, Al-Ahsa oasis of eastern Saudi Arabia. Measurements of Yemeni bee were smaller than Carniolan bee for body weight, head structures, including antenna, flagellum, and proboscis length, thorax appendages, including femur length, tibia length and width, metatarsus length and width of the right hind leg, and length and width of the right forewing and hind wing, abdominal characteristics, including the length of the 3rd and 4th abdominal tergites and sternites, and length and width of the 1st and 4th wax mirrors. It could be concluded that with the exception of the number of hamuli, worker Yemeni bee body size and morphometric parameters related to the colony productivity were smaller than Carniolan bees under environmental conditions of the study region.

Exploring the non-coding regions in the mtDNA of some honey bee species and subspecies.

The sequence of the DNA contains coding and non-coding regions. The role of the non-coding regions is not known and is hypothesized to maintain the structure of the DNA. This study aimed to investigate the structure of the non-coding sequences in honey bees utilizing bioinformatics. The non-coding sequences of the mtDNA of three honey bee species Apis dorosata, Apis florea, Apis cerana, and ten subspecies of Apis mellifera were investigated. Different techniques were utilized to explore the non-coding regions of these bees including sequence analysis, phylogenetic relationships, enzymatic digestion, and statistical tests. Variations in size and sequences of nucleotides were detected in the studied species and subspecies, but with the same nucleotide abundance (i.e. nucleotides A were more than T and nucleotides G were less than C). The phylogenetic tree based on the non-coding regions was partially similar to the known phylogenetic relationships between these bees. The enzymatic digestion using four restriction enzymes confirmed the results of the phylogenetic relationships. The statistical analysis based on numerical codes for nucleotides showed the absence of significant variations between the studied bees in their sequences in a similar way to results of neutrality tests. This study suggests that the non-coding regions have the same functional role in all the studied bees regardless of the number of nucleotides, and not just to maintain the structure of the DNA. This is approximately the first study to shade lights on the non-coding regions of the mtDNA of honey bees.

Repellent and Toxicant Effects of Eight Essential Oils against the Red Flour Beetle, Tribolium castaneum Herbst (Coleoptera: Tenebrionidae).

This study was conducted to compare the repellent effect and contact toxicity of eight essential oils (EOs), including Syzygium aromaticum, Allium sativum, Eucalyptus camaldulensis, Lavandula officinalis, Simmondsia chinensis, Matricaria chamomilla, Citrus limon, and Prunus dulcis, against adults of Tribolium castaneum Herbst. Four concentrations (1, 5, 10, and 15% in acetone solvent) of each EO were tested. The 5, 10, and 15% concentrations of S. aromaticum EO had a high repellency effect against T. castaneum compared with A. sativum, E. camaldulensis, L. officinalis, S. chinensis, M. chamomilla, C. limon, and P. dulcis after 30 min of exposure. The repellency test of the S. aromaticum, E. camaldulensis, L. officinalis, M. chamomilla, C. limon, and P. dulcis EOs on T. castaneum has shown that the mortality percentages enhanced with the increase in the EOs concentration and also with the exposure time. The 15% concentration of P. dulcis and M. chamomilla EOs have a significant impact on the mortality rate of T. castaneum compared with S. aromaticum, A. sativum, E. camaldulensis, L. officinalis, and S. chinensis after the 24 h of contact test. Moreover, the 15% concentration of the C. limon EO caused a greater mortality percentage compared with S. aromaticum, A. sativum, E. camaldulensis, and L. officinalis. It could be concluded that using the S. aromaticum EO as a repellent oil and using P. dulcis, M. chamomilla, and C. limon for contact toxicity to treat the flour infested by T. castaneum can play an important role in protecting stored grains and their products.

Post grafting time significantly influences royal jelly yield and content of macro and trace elements.

Royal jelly (RJ) is commercially harvested after the 4th day of queen larval age. In the current study, it was harvested after 24, 48, 72, and 96 hours after grafting of 1-day larval age queens to investigate changes in macro and trace elements associated with harvesting time. The RJ yields were significantly affected by harvest time, and the highest yield was obtained 72 hours after grafting. The highest phosphorus (P) and zinc (Zn) contents were obtained from RJ harvested 24 hours after grafting. Royal jelly harvested 48 hours after grafting had the highest concentrations of magnesium (Mg), calcium (Ca), potassium (K), sodium (Na), iron (Fe), and manganese (Mn). Likewise, RJ harvested 96 hours after grafting had higher concentrations of copper (Cu). Royal jelly harvested 72 hours after grafting showed the second rank for P, Mg, Ca, K, Na, Fe, Cu, and Mn concentrations. In descending order, P, Mg, Ca, and K were the most dominant elements in RJ harvested at different times after grafting. The Mg, Ca, K, Na, Cu, and Mn concentrations in RJ were all positively correlated, and P, Fe, and Zn were positively correlated. The P and Zn were negatively correlated with Ca, Cu, and Mn. It was concluded that macro and trace element contents in RJ can differ depending on the harvest time after grafting. We recommend harvesting RJ at 72 hours after grafting for possible use as healthy nutritional human food supplement.

Macro- and trace elements content in honeybee pollen loads in relation to the harvest season.

The content of macro-and trace elements in honeybee pollen loads collected from the Al-Ahsa oasis, Saudi Arabia during spring, summer, autumn, and winter in 2018/2019 were determined. After critical screening and observation, we found that the major pollen floral resources were rapeseed, sunflower, summer squash, and date palm. The highest values of sodium (Na), potassium (K), calcium (Ca), magnesium (Mg), phosphorus (P) and manganese (Mn) were found in pollen loads trapped during spring and winter. The highest levels of copper (Cu) and iron (Fe) were found from pollen loads harvested during autumn, while the highest content of zinc (Zn) was found in pollen loads collected during the summer. Higher concentrations of the essential elements (Na, K, Ca, Mg and P) for human nutrition found in pollen loads collected during the spring and winter seasons, make them valuable food supplements. Finally, the elements contained in pollen loads can be related to the season of pollen load collection.

The relationship between comb age and performance of honey bee (Apis mellifera) colonies.

A study on the relationship between the age of comb and the activity of the hybrid Carniolan honey bee colonies in collecting pollen activity, worker brood production, colony strength, and honey yield was conducted. In comparison to colonies with combs aged 4-years, colonies with combs aged 1, 2 and 3-years significantly exceeded in the number returning workers, number returning workers with pollen loads, rate of storing pollen, rate of worker brood production, and size of colony population. Colonies with combs aged 1, 2 and 3-years produced significantly more honey than colonies with combs aged 4-years (5.25, 4.90 and 4.65 kg/colony vs. 4.45 kg/colony, respectively). It can be concluded that the foraging rate, gathering and storing pollen, brood production, colony population size, and honey yield significantly depended on the age of combs. Beekeepers can replace old combs with new ones to increase brood and honey production.

Protein content and amino acids composition of bee-pollens from major floral sources in Al-Ahsa, eastern Saudi Arabia.

Protein content and amino acids composition of bee-pollens from major pollen floral sources in Al-Ahsa, Saudi Arabia were determined to investigate the nutritive value of pollen protein relative to requirements of honeybees and adult humans. The major pollen sources were alfalfa, date palm, rape, summer squash, and sunflower. Bee-pollens from alfalfa and date palm showed high content of crude protein and amino acid concentrations. Bee-pollen from sunflower had low content of those components. Eighteen amino acids were found in bee-pollens from the five major floral sources. The highest concentrations of individual amino acids valine, leucine, isoleucine, phenylalanine and proline were obtained from alfalfa bee-pollen; lysine, arginine, cysteine, tryptophan and tyrosine from date palm; methionine, histidine, glycine and alanine from summer squash; threonine, serine and glutamic acid from sunflower; and aspartic acid from rape bee-pollen. The amino acid composition obtained from sunflower bee-pollen showed the lowest concentrations of the essential amino acids: isoleucine, leucine, methionine, phenylalanine and valine. Apart from methionine, arginine and isoleucine, the essential amino acids of bee-pollen from alfalfa, date palm, summer squash and rape exceeded the honeybees' requirements. Methionine was the limiting amino acid in bee-pollens from the five selected sources. Concentrations of essential amino acids in the tested bee-pollens were variable and significantly correlated to their botanical origin of pollen. Bee-pollens from alfalfa, date palm and summer squash was found to be rich source of protein and amino acids for bees and for humans.

Nectar and pollen sources for honeybees in Kafrelsheikh province of northern Egypt.

This study was conducted at the apiary of the Beekeeping Research Section at the Sakha Agricultural Research Station, ARC, Kafrelsheikh, and other apiaries in Kafrelsheikh province, during two successive years 2015 and 2016. The study aimed to survey nectar and pollen floral resources in Kafrelsheikh province. Ninty seven plant species belonging to 33 families were recorded as nectar sources, and 82 plant species belonging to 36 families were recorded as pollen sources during the whole year. The largest amount of monthly trapped pollen was obtained during May followed by August. It can be concluded that, beekeepers in Kafrelsheikh province can harvest good honey yield at the end of blooming seasons of citrus (Citrus spp.) during March and April, Egyptian clover (Trifolium alexandrinum L.) during May and June, loofah (Luffa aegyptiaca Mill.) during June to October, cotton (Gossypium spp.) during July and August, and banana (Musa spp.) during August and September. They also, could be trapping pollen loads collected from faba bean (Vicia faba L.) and flax (Linum usitatissimum L.) during January to March, date palm (Phoenix dactylifera L.) during March and April, Egyptian clover during May and June, summer seed watermelon (Citrullus lanatus var. colothynthoides L.) during June and July, loofah and maize (Zea mays L.) during June to November.

Comparison of the activity and productivity of Carniolan (Apis mellifera carnica Pollmann) and Yemeni (Apis mellifera jemenitica Ruttner) subspecies under environmental conditions of the Al-Ahsa oasis of eastern Saudi Arabia.

This study was conducted at the apiary of the Agricultural and Veterinary Training and Research Station of King Faisal University in the Al-Ahsa oasis of eastern Saudi Arabia. We performed a comparison between Carniolan (Apis mellifera carnica Pollmann) and Yemeni (Apis mellifera jemenitica Ruttner) honeybee races to determine the monthly fluctuations in foraging activity, pollen collection, colony growth and honey yield production under the environmental conditions of the Al-Ahsa oasis of eastern Saudi Arabia. We found three peaks in the flight activity of the two races, and the largest peaks occurred during September and October. Compared to Carniolan bee colonies, the performance of Yemeni bee colonies was superior in terms of stored pollen, worker and drone brood rearing, and the adult population size. The Carniolan bee colonies produced 27.77% and 27.50% more honey than the Yemeni bee colonies during the flow seasons of alfalfa and sidir, respectively, with an average increase of 27.64%. It could be concluded that the race of bees is an important factor affecting the activity and productivity of honeybee colonies. The Yemeni bee race produced more pollen, a larger brood and more bees, which exhibited a longer survival. The imported Carniolan bees can be reared in eastern Saudi Arabia, but the Yemeni bee race is still better.