الفهرس 
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Abstract
SUMMARY
The current study was carried out in the apiary and laboratory of Department of Bee Research Quanater, Qaluobia Governorate, Plant Protection Institute and Department of Pathology, Animal Health Research Institute, Agricultural Research Centre Giza governorate to study some biological aspects and recent trends for controlling wax worms in Egyptian apiaries. The following experiment were carried out:
1- Identification and manifestation of infestation and control wax moth via questionnaire form, which distributed on beekeepers in different parts of Egypt, to identify the knowledge and skills levels for damage and control wax moth in Egypt.
2- Efficiency evaluation of some feeding materials for rearing greater wax moth (Galleria mellonella L).
3- Efficiency evaluation of methanolic extracts of some plants with different concentrations against greater wax moth (Galleria mellonella L.) and evaluation the efficiency of aqueous extract of some plants on greater wax moth and adult honey bee workers.
4- Efficiency evaluation of some essential oils (cinnamon, garlic, clove, camphor, and menthol oils) against greater wax moth (Galleria mellonella L.) and adult honey bee workers.
5- Toxicity evaluation of nanoparticle compounds (Raval and Chetozan) against greater wax moth larvae (Galleria mellonella L.) and adult honeybee workers.
6- Preservation of bee wax combs from wax moth infestation in honeybee colonies in the apiary by using nanoparticle compounds (Raval and Chitozan)
7- Histological studies of mid gut of greater wax moth larvae (Galleria mellonella L.) treated with nanoparticles compounds (Raval and Chitozan) and water extract of some plants.
6.1. Questionnaire form for identification, manifestations of infestation and control of wax worms.
To detect the knowledge and skills level for identification, manifestation and control methods of wax moth, 50 forms were distributed randomly on Egyptian beekeepers during their gathering through workshops, symposiums and Egyptian honey exhibitions. The results revealed that the identification of questionnaire recipients of 50 apiaries in different governorates in Egypt, showed that 98% of apiaries in questioner owned by beekeepers themselves, 44% had 100-1000 colony, 62% of them had built up their apiaries from years between 2000-2020, 100% of them product honey and 100% of them use honeybee race, (Apis mellifera carnica) and their hybrid beside other honeybee strains. Satisfaction with work in the field of beekeeping (98% of beekeepers prefer their children to work in beekeeping, 22% of beekeepers always completely satisfied with their income from the apiary, 60 % of them scarcely think that beekeeping in recent years has become unrewarding and they are thinking of another projected, 78% indicated that sometimes beekeeping is difficult and expensive).
For bee diseases: data showed that for first disease, Varroa disease, 56% of beekeepers indicated that degree of spread of disease was medium, 66% detect medium high degree of Nosema. The brood diseases were Chalk brood, Stone brood and American foulbrood diseases, 6% of beekeepers indicated that degree of spread of diseases was medium. Bee pests problems: Wax worms, 46% of beekeepers indicated that the degree of spread of wax worms was medium; Vespa orintalis, about 48% indicated that the degree of spread of this enemy was medium in their apiaries otherwise 0% do not know; Plistes gallicas, 10% detect high degree of Plistes gallica, 36% medium, Braula coeca pest, 0% detect high degree of disease, where as 14% medium, 42% few, meanwhile 28% detect no percent of disease, at last 16% do not know. At last, Acerontia atrops, 0% told that it was high, 58% few, while 26% say there is no Acerontia atrops in their colonies, otherwise 6% do not know.
For animal and Bird pests’ problems: The bee eater, 24% of beekeepers indicated that the degree of spread of pest was high, 24% medium. The Rats, 38% told that it was medium in their colonies.
Identification of wax worms, symptoms of infestation and damage to wax and methods of prevention and control of wax worms, The questionnaire showed that 100% of beekeepers have knowledge about wax moth, and it belong insects, the harmful phase of the insect wax moth is larvae (worm) and wax moth is an insect that infects wax combs. The 100% of beekeepers know that manifestations of wax worm infestation is the presence of wax worm butterflies in the beehives and on the combs in the store. Also, the presence of silk threads in the apiary, presence of larvae on the infected combs inside the beehive in the store, presence of damaged wax combs and the remains of wax inside the beehive. 100% of beekeepers to prevent wax worms keep bee colonies strong, clean and not throw wax residues in the land of apiary. To control wax worms on the combs stored inside the store100% of beekeepers using Micronized sulfur, Paradex.
6.2. Efficiency evaluation of some feeding materials for rearing greater wax moths (Galleria mellonella L.) and their effect on larval stage (day), larval weight (g), (%) pupation and adult emergences (%).
Larvae of the greater wax moth (Galleria mellonella L.) were collected from naturally infested honeybee colonies. Wood boxes 40 x 30 x 30 cm were used for rearing process under laboratory, condition with 25 ± 5ºC and 70 ± 5% relative Humidity. Four food regimes (media): i.e. artificial diet, Two-years old bee wax; one-year old wax and wax foundation were evaluated in this study, the experiment extended for six weeks, at the end of the experiment, larval age in each and larval stage (day), larval weight (gm), % of pupation and adult emergences of wax moth larvae were estimated. The results concluded that, the highest larval stage was recorded in case of artificial diet and the lowest was found with wax foundation where the average of larval stage was 52.33, 47.33, 41.00 and 15.66 day for artificial diet, two -years old wax combs, one-year old wax combs and wax foundation, respectively. Significant differences were found for larval stage among feeding materials used. For larval weight (gm), the same trend was obtained, the highest larval weight (gm) was recorded with artificial diet and the lowest larval weight was found in wax foundation. The average larvae weight was 0.652, 0.588, 0.282 and 0.039 mg/larva for artificial diet, two -years old combs, one-year wax combs and wax foundation, respectively. Significant difference was found between all media and rearing materials tested. For percentage of pupation (%), it recorded 100% with all materials used, but no adult emergences were observed in case of wax foundation, meanwhile, it was 90% with artificial diet, it increased to rich 100% when two-year old combs and one-year old wax combs were used.
To sum up, from the previous results, it recommended old bee wax for mass rearing of wax moth, when needed for scientific experiments.
6.3 Management greater wax moth (Galleria mellonella L.)
6.3.1 Efficiency evaluation of methanolic extract of some plants against greater wax moth larvae (Galleria mellonella L.)
The current work was carried out in the laboratory of Department of Bee Research, Quanater, Qaluobia Governorate to evaluate the toxicity of methanolic extract of four plants against greater wax moth larvae (Galleria mellonella L.). Different concentrations of methanolic extract (0.25, 0.50, and 1.00 mg/1ml) of bestachia (Adhatoda vasica); dumb cane (Diffenbachia bicta); the worm wood santonica (Artemisia sp.) and Thyme (Thymus sp.) were evaluated. The percentage of mortality was 26.66, 20.00, 0.00 and 0.00% when used 0.25 mg/1ml water of dumbcane, bestachia, santonica and thyme, respectively and in the case of using .050 mg/1ml the percentage of mortality was 33.30, 26.66, 13.30 and 6.66% for dumb cane, bestachia, santonica and thyme, respectively. Finally the percentage of mortaliy for 1.00 mg/1ml was 40.00, 33.30, 20.00 and 13.30% for dumb cane, bestachia, santonica and thyme, respectively. The results also revealed that no significant differences were found between bestachia and dumb cane extract. Regarding the concentration, the highest percentage of mortality of garter wax moth larvae was recorded in case dumb cane, bestachia followed by thyme, meanwhile, santonica extract did not affect the larvae mortality where thier percentage of mortality was 13.33 at 1.00 mg/1ml concentrate.
from the previous results, it recommended using methanolic extract of dumb cane, bestachia and thyme for controlling greater wax moth larvae (Galleria mellonella L.) that gave promising results against them.
6.3.2 Efficiency evaluation of aqueous extract of some plants against 2nd-inster larvae of greater wax moth(Galleria mellonella L.).
This evaluation was carried out in the laboratory of Department of Bee Research Quanater, Qaluobia Governorate. The filtered of aqueous extract of some plants (dumb cane, bestachia, santonica and thyme) was evaluated against greater wax moth (Galleria mellonella L.) directly by added them separately to 10gm of old bee wax. Results indicated that the water extract of tested material was not effect on greater wax moth larvae during the first three days of treatment under laboratory condition. They were more effective after 7 days, where the accumulative mortality (%) of 2nd instar larvae of greater wax moth (Galleria mellonella L.) was 63.33, 20.00, 96.60. 39.99 and 0.00 (%). It was increased to rich 79.93, 50.00, 96.60, 49.99 and 0.00 (%) after 14 days for bestachia, dumb cane, thyme, santonnica and untreated check (control), respectively. The highest number of emerging wax moth was recoded in case untreated check (control) group 30.00 adult with 100% of adult emerging as compared with 20.00, 30.00. 3.33 and 30.00 (%) for bestachia, dumb cane, thyme and santonnica extract respectively.
To sum up, aqueous plant extract of bestachia, dumb cane, thyme and santonica gave moderate results against greater wax moth.
6.3.3 Evaluation of the toxicity of aqueous extract of some plants against adult honeybee workers.
To study the evaluation of aqueous extract of bestachia, dumb cane, thyme and santonnica against adult honey bee workers fifty adult honeybee workers were used for this assay, they were transferred into especial cages, and each cage contained 50 of adult honeybee workers and three cages were used for each plant (replicates). The adult honey bee workers were treated with aqueous extract of tested plants by spring them directly. The data clearly showed that the accumulative corrected mortality (%) of adult honeybee workers after 24 hrs. of treatment was 0.00, 6.16, 0.00, and 4.08(%) but it was 8.63, 19. 72, 2.04 and 7.47(%), after 48hrs. at the end of the bioassay after 72 hrs. it was 8.83, 26.53, 8.16 and 13.60 (%) for bestachia, dumb cane, thyme and santonnica, respectively. The percentage of mortality clearly showed that dumb cane was more toxic for adult workers of honey bees, flowed by santonica; mean while bestachia and thyme extract were less toxic for adult honeybee workers.
from the obtained results, it can be concluded that, aqueous extract of bestachia, dumb cane, thyme and santonnica is highly save for adult honey bee workers as compared with untreated check group.
6.3.4. Efficiency evaluation of some essential oils against greater wax moth larvae (Galleria mellonella L.) and adult honey bee workers.
The assay was performed on fourth instar larvae of wax moth, 10 larvae were used for each (replicates). Three replicates were used for each essential oil. The experiment were carried out under the laboratory condition in Department of Bee Research Quanater, Qaluobia Governorate condition to study the toxicty of some essential oils; garlic (Allium sativum) camphor, (Cinnamomum camphor), menthol (Mentha sp.) cinnamon, (Cinnamomum verum) and clove (Dianthus sp.)against greater wax moth larvae. The data concluded that, cinnamon, garlic, menthol, clove, and camphor oils were highly effective against greater wax moth larvae, where hundred percentage mortality was attained after 48 and 96 hrs. of treatment with cinnamon and garlic oil. For clove and mint oil, they achieved 100% mortality after 120 hrs. of treatment meanwhile camphor oil provided 96.60% accumulative mortality after 120 hrs. Moreover, all the essential oils evaluated were highly safe for adult honeybee workers under laboratory conditions. The rate of accumulative corrected mortality after 72 hrs. for adult honey bee workers was 15.96, 9.02, 13.18 , 29.16 and 8.88% for cinnamon, clove, camphor, mint, and garlic oils, respectively.
To sum up, from the aforementioned results we can recommended that bee keepers can use cinnamon, garlic, menthol, clove and camphor oil for controlling greater wax moth larvae that provided high percentage of mortality in the same time, they highly save for adult honey bee workers.
6.3.5. Toxicity evaluation of nanoparticle compound (Raval and Chetozan) against greater wax moth larvae (Galleria mellonella L.) and adult honeybee workers.
To study the toxicity evaluation of nanoparticle compounds (Raval and Chitozan) against greater wax moth larvae (Galleria mellonella L.), twenty-seven plastic containers with perforated lids were prepared for this study, each prepared with 10 gm of old bees wax. Ten 2nd instar larvae of greater wax moth (Galleria mellonella L.) were introduced in each container for assay. Three replicates of each tested material of nanoparticle compounds (Raval and Chetozan) with concentration (10, 20, 30 and 40%) were evaluated. The obtained data revealed that, the number of dead larvae increased by increasing the concentration of tested compounds. For Raval compound, all exposed larvae were dead after 120 hrs. of treatment with all tested concentrations. It also found that Chetozan compound was more toxic against greater wax moth larvae more than Raval, compound with 40 and 30% concentration, where all the exposed larvae were dead after 96 hrs. of treatment but they dead after 120 hrs. with 20 and 10 (%) concentration. The results also clearly summarized that the tow nanoparticles compounds were highly save for adult honey bees workers, where the percentage of mortality was 4.60 and 6.00% for Raval and Chetozan compound with 40% concentration, respectively.
To sum up nanoparticle compounds (Raval and chitozan) in highly recommended for controlling greater wax moth (Galleria mellonella L.) larvae, where they highly toxic for wax moth worms and highly save for honey bees.
6.3.6. Preservation of bee wax combs from wax moth infestation in honeybee colonies in the apiary by using nanoparticle compounds (Raval and Chetozan).
The current study was conducted in the apiary belong to Department of Bee Research Quanater, Qaluobia Governorate to evaluate nanoparticle compounds (Raval and Chetozan) in preservation of empty bee wax combs from wax moth infestation. Nine boxes of beehive were prepared for this study, each contained four empty bee wax combs and three boxes were used for each compound (replicates). from the obtained results it can be concluded that the nanoparticles compounds (Raval and Chitozan) provided some protect for bee wax combs as compared with untreated check (control). At end of the experiment, the total larvae and pupa of greater wax moth (Galleria mellonella L.) were zero and the adult of wax moth was 18, 33 and 193.00 adult with an average of 6.00,11.00 and 64.33 adult/bee hive for Raval, Chetozan and untreated check (control), respectively
6.3.7. Histological studies of mid gut of greater wax moth larvae (Galleria mellonella L.) treated with nanoparticles compounds (Raval and chitozan) and aqueous extracts of some plants.
To study the effect of nanoparticle compounds (Raval and chitosan) and aqueous extract of some plants (dumb cane, bestachia, santonica and thyme), greater wax moth larvae were treated with aforementioned compounds. After treatment, the dead greater wax moth larvae (Galleria mellonella L.) were stored in 10% formalin solution for preservation until dissection process and for glass slides preparation. Dead larvae were dissected and the midguts of larvae were excited and preserved in 10% neutral buffer formalin this study was carried out in Department of Pathology, Animal Health Research, agricultural centre, Giza gafearnmeernm. The results of histological studies clearly showed that, certain histological changes through cross section of mid gut of six instar greater wax mouth larvae. Treatments were detected that the cytoplasmic extrusion appeared as the apical margin of cells as confluent mass and muscular layers were broken in some parts, large amount of secretions released in the lumen of the mid gut while a few amount was attached to the apical margin cell. Much destruction of the mid gut took place and large number of epithelial cells becomes vacuolated and the cytoplasm appeared as confluent masses because of the hydropic analysis of the epithelium. Sever vacuolar degeneration with elongation and necrosis of the epithelial cell lining.