Sperm quality of drones of Apis mellifera L. selected for royal jelly production

Royal jelly–induced asthma and anaphylaxis: Clinical characteristics and immunologic correlations

This investigation explored the effects of royal jelly (RJ) administration in rams on reproductive parameters, with a specific focus on semen quality, post-thaw characteristics, sperm function, and testicular health. Rahmani Rams (n=15) were divided into 3 groups: the control group received 1 mL of distilled water orally (RJ0), and two treatment groups received 250 mg (RJ250) or 500 mg (RJ500) of RJ three days per week, every other day per head for 6 months. Oral administration of RJ resulted in significant improvements in sperm mass motility, viability, and plasma membrane integrity (p < 0.001). Conversely, ejaculate volume, sperm concentration, and the percentage of morphological abnormalities showed no significant differences between groups (p > 0.05). The RJ administration resulted in a significant reduction in oxidative markers such as MDA, H 2 O 2 , and NO, and an increase in TAC and CAT compared to the RJ0 group (p <0.01) in raw and post-thawed semen. In cryopreserved semen, the progressive motility, viability, and membrane function were significantly higher in all RJ-treated rams (p <0.01). The majority of sperm parameters, including progressive motility, distance average path, distance curve linear, distance straight line, average path velocity, curve linear velocity, and straight line velocity, were superior in the RJ groups compared to the untreated ram group (p <0.05). The orally administered RJ affected both sperm membrane and acrosome status significantly (p <0.01). Specifically, RJ supplementation significantly increased the proportion of viable spermatozoa with intact acrosomes, while concurrently reducing the percentages of both viable acrosome-damaged and non-viable acrosome-intact cells across all treatment groups (p <0.05). Sperm viability was significantly higher in the RJ250 and RJ500 groups compared to the RJ0 treatment (p < 0.001). Furthermore, RJ administration significantly decreased the incidence of early apoptotic, late apoptotic, and necrotic spermatozoa relative to the RJ0 group (p < 0.05). Regarding the microbial profile, both coliform and total bacterial counts in post-thawed semen were significantly lower in RJ-treated rams than in the RJ0 group (p < 0.05). Oral administration of RJ significantly preserved testicular histoarchitecture, characterized by the maintenance of seminiferous tubule integrity, an increased population of spermatogonia, and the distinct presence of intraluminal spermatozoa. Furthermore, RJ treatment maintained the critical ultrastructural components of the spermatozoa, including the structural integrity of the plasma membrane, the mitochondrial sheath, and the nucleus. Collectively, these findings suggest that oral RJ supplementation enhances reproductive performance and post-thaw semen quality in rams, likely mediated by its potent antioxidant properties and its protective role during the cryopreservation process.

The aim of this study was to investigate the ability of royal jelly in protecting rat testes against carbon tetrachloride induced testicular damage in male Wistar rats. Thirty six rats (weighting 220-235 g) were divided into six groups. The first group administrated distilled water and basal diet and kept as negative control, the second group administrated CCl4 as in equal volume with olive oil (1/1) and kept as negative control, the third group and fourth group were administrated royal jelly at two doses 150 mg/kg and 300 mg/kg , the fifthgroup and In sixth group were administrated CCl4 with royal jelly at two doses doses 150 mg/kg and 300 mg/kg respectively. At the end of experiment (50 days) the results show significant decrease in the sperm concentration, sperm viability, sperm motility in test sand significant increase in the sperm abnormality in the group of rats treated with CCl4 only while the results appear significant increase in sperm concentration, sperm viability, sperm motility in testes and significant decrease in the sperm abnormality in the groups of rats treated with CCl4and royal jelly at two doses respectively but all the sperm parameters like negative control in the group of rats treated with royal jelly at two doses only in compare with negative control. As well as the study demonstrated a significant decrease in sex hormones levels (T, LH,FSH)in the group of rats treated with CCl4 only while the results appear significant increase in the levels of six hormones in the group of rats treated with CCl4 and royal jelly at two doses respectively but all the levels of six hormones as negative control in the groups of rats treated with royal jelly at two doses only in compare with negative control.The histological in testes show complete swallowing of seminiferous tubules with degenerated germ cells and hypoplasia of Leydig cellsbut showing mild degeneration in spermatogenic cells and moderate degeneration of leydig cells in the groups of rats treated with CCl4 and royal also the histological results show normal seminiferous tubules with complete spermatogenic cells in the groups of rats treated with royal jelly only at two doses only. In conclusion, royal jelly is nonviolent and active in the treatment of infertility problem in male

Background: Cisplatin is one of the cytotoxic agents in the treatment of cancer and has side effects on sperm function. Objective: This study aims to investigate the ameliorative role of dietary bee honey and royal jelly against cisplatin-induced alterations in sperm function parameters in male wistar albino rat. Methods: Male wistar albino rats of same age and weight were randomly divided into four groups; G, I: control group which was given 0.9% saline, G: II: cisplatin (7 mg/kg/d) was injected intraperitoneally for 15 d, G, III bee honey with royal jelly (500 mg/kg/d of honey and 100 mg/kg/d of royal jelly) fed orally daily for 15 d, G, IV: cisplatin (7 mg/kg/d) was injected intraperitoneally and honey (500 mg/kg/d) and royal Jelly (100 mg/kg/d) fed orally daily for 15 d. The sperm function parameters like sperm count, sperm viability, sperm motility, weight of epididymis and testis were measured by using a hemocytometer under the microscope at 100X magnification. Results: Cisplatin treated rats revealed a significant decrease in sperm count, sperm viability, sperm motility, weight of epididymis and testis as compared to control group. Royal jelly and honey treated group of rats revealed a significant increase in all sperm function parameters compared to control group. Dietary bee honey with royal jelly along with cisplatin-treated rats revealed significant increase as compared to animals treated with cisplatin (G, II). Conclusion: Bee honey and royal jelly could be used as dietary preventive natural products against cisplatin induced sperm function parameters alterations during the treatment of cancer.

ABSTRACTIn trying to protect honey bee species and maintain genetic diversity, employing effective approaches for drone sperm conservation is crucial. Based on literature, drone sperm cryopreservation extenders and methods have not been fully optimized. Our research aim was to enhance drone bee sperm preservation by incorporating royal jelly (RJ) in the extender of the cryoprotectant‐free vitrification method. Drone sperm was collected from adult drone bees (36 days old) using the manual inversion method. Different concentrations of RJ (0%, 0.5%, 1%, 2.5% and 5%) were added to the extender. Diluted sperm were cryopreserved using a cryoprotectant‐free vitrification technique where 10 µL aliquots of the diluted sperm were directly dropped into the liquid nitrogen and then stored. Data were analysed based on a completely randomized design with ten replications. Sperm quality parameters, including motility, viability and DNA damage, were evaluated in vitro. Queens were artificially inseminated to measure the ability of motile sperm to reach the spermathecae. The results showed that 1% and 2.5% of RJ supplementation significantly enhanced sperm motility and viability and reduced DNA fragmentation compared to control and higher RJ concentrations. Specifically, the 1% RJ group resulted in the highest sperm viability, while both the 1% and 2.5% groups maintained lower DNA fragmentation rates. Queens inseminated with sperm treated with 1% and 2.5% RJ showed a notably higher number of motile sperm in their spermathecae. In conclusion, supplementation of 1% RJ to the cryoprotectant‐free vitrification media may improve drone sperm quality parameters post‐warming. Our findings provide valuable insights into optimizing drone bee sperm preservation, contributing to the conservation of these vital pollinators.

Background: Bleomycin (BL) is a glycopeptide antibiotic obtained from the bacterium Streptomyces verticillus which is routinely used for treatment of human cancers. Royal jelly (RJ) is a production from the hypo pharyngeal, mandibular and post cerebral glands of nurse bees. RJ consists of 66% water, 15% sugars, 5% lipids, and 13% proteins, essential amino acids and vitamins.Objective: The aim of present study was to evaluate protective effect of royal jelly on sperm parameters and malondialdehyde (MDA) production in rat.Materials and Methods: Forty adult male wistar rats (220±20gr) were randomly divided into 4 groups (n=10). Control group (CG) received normal saline 10 ml/kg twice a week with Intraperitoneal (I.P) for 48 days (0.3 ml/rat(. Royal Jelly group (RJG) received jelly (100 mg/kg daily) for 48 days orally. Bleomycin group (BLG) received BL (10 mg/kg twice a week) with I.P for 48 days. Royal Jelly+ Bleomycin group (RJ+BLG) received royal Jelly (100 mg/kg /day) orally concomitant with BL administration. Sperm count, motility, and viability were investigated and chromatin quality and DNA integrity were also analyzed. Serum testosterone and MDA concentrations were measured as well.Results: BL caused decline significantly (p<0.05) sperm count, sperm viability, motility as well as testosterone concentration compared to control group while significant (p<0.05) increases in immature sperm, sperm with damaged DNA and MDA concentration were announced in BL in comparison with CG and RJ+BLG. Royal jelly improved Bleomycin-induced toxicity on sperm parameters and testosterone and MDA concentrations. Conclusion: The present results support the idea that BL adversely affects sperm parameters and MDA and the RJ with antioxidant properties has positive effects on these parameters.This article extracted from M.Sc. thesis. (Tayebeh amirshahi)

The current study was carried out to investigate the protective effects of royal jelly supplementation on sperm motility, viability and pH value during the liquid storage of boar semen at 16 °C and 4 °C, at various periods of time (0, 24, 48, 72 and 96 h). Semen samples were collected from 11 boars, diluted with a long-term extender and supplemented with different concentration of royal jelly (0%, 0.5%, 1% and 2%) at a final concentration of 50 × 106sperm/ml. In the laboratory, the semen was assessed for sperm morphology, viability (eosin-nigrosin staining), subjective motility and objective sperm motility by sperm class analyzer. In total, 396 tests for sperm viability and motility were performed. The longer storage time and the lower incubation temperature showed lower sperm motility and viability results. The results showed that royal jelly supplementation at 1% concentrations protected the functionality of the sperm plasma membrane during the liquid storage time of 96 h at 16 °C. Sperm subjective and objective motility results in samples stored at 4 °C decreased with higher royal jelly concentrations and longer storage time, and differ significantly from the results in samples stored at 16 °C (P&lt; 0.05). Our data showed that royal jelly supplementation at lower concentrations can improve boar semen motility and viability parameters during liquid storage at 16 °C for 96 h.

Royal jelly supplementation in semen extender enhances post-thaw quality and fertility of Nili-Ravi buffalo bull sperm

This study investigates the impact of bee pollen nutrition on the royal jelly production of honey bees (Apis mellifera). Results demonstrate that pollen diet significantly impacts hypopharyngeal gland (HPG) development and the expression of genes associated with royal jelly biosynthesis. Bees fed Brassica napus pollen exhibited superior HPG development, and increased mrjp1 expression (encoding a key royal jelly protein). While the cyp450 6AS8 gene expression (encoding a key enzyme in 10-HDA biosynthesis) was increased by pollen consumption, no distinct expression patterns were observed among the different pollen types tested. An in vitro bee cage platform for royal jelly production has been established to further understand the mechanisms behind royal jelly production in bees. The experiment demonstrated a positive correlation between the number of worker bees and the total yield of royal jelly per cage. However, when the number of worker bees is low, the amount of royal jelly each individual worker bee needs to produce increases. In conclusion, these findings enhance our understanding of the role of bee pollen nutrition in royal jelly production. Furthermore, the results from this in vitro bee cage platform suggest that the number of worker bees is a critical factor in royal jelly production, and that bees may possess a controllable mechanism for regulating royal jelly secretion.

The present work was designed to study the effect of propolis (P) with or without royal jelly (RJ) and bee honey (H) on age of sexual puberty, semen quality and fertility of New Zealand White (NZW) rabbit males. Sixty pre-pubertal NZW rabbit males were used in the present study and were randomly divided into 3 groups (20 bucks per group); males were administered orally with 0.5 mL of a solution/ kg body weight (BW), 3 times weekly for 6 weeks. The solution contained: 1) water for control (1st group), 2) 15 mg propolis with 200 mg royal jelly (RJ) suspended in 0.25 mL bee honey (H) + 0.25 mL water (2nd group), 3) 15 mg propolis (P) was suspended in 0.5 mL water (3rd group). Fertility of rabbit bucks was tested using artificial insemination of 60 hybrid nonparous rabbit females (20 females per each experimental group). Male NZW rabbits received propolis with or without royal jelly and bee honey showed significant (P<0.05) earlier age at puberty (earlier ages at testis descending into scrotum, separation of penis from sheath, fighting, first ejaculation, appearance of sperm in testis and epididymis). The results also revealed significant (P<0.05) increase of ejaculate volume, percentage of sperm progressive motility, sperm-cell concentration and seminal plasma fructose concentration in treated groups compared to control group. On the other hand, reaction time, percentages of dead and abnormal spermatozoa increased significantly (P<0.05) for control group compared to the other two groups. Blood plasma concentration of testosterone increased significantly (P<0.05) for both 2nd and 3rd groups in comparison with control. Otherwise, plasma concentrations of cholesterol, aspartate amino transferase (AST) and alanine amino transferase (ALT) were reduced significantly (P<0.05) for 2nd and 3rd groups compared to control group. Rabbit bucks treated with either propolis with or without royal jelly and bee honey showed better fertility (higher conception rate and litter size) than control bucks. In conclusion, the results of the present study showed that oral administration of NZW rabbit males with propolis only or with royal jelly and bee honey could be used beneficially to have earlier age of puberty, improve semen quality and fertility. This improvement was also mirrored on better liver functions.

Multi-organ proteome reveals different nursing ability between two honeybee srocks.

To alleviate the deleterious effect of heat stress during summer conditions on male rabbits’ reproduction, 40 V Line adult rabbit bucks (on av. 8 mo old) were divided into 4 experimental groups and exposed to temperatures ranging from 23 to 36°C. Bucks in the 1st, 2nd, 3rd, and 4th group were supplemented with 0, 50, 100 or 150 mg of Chinese royal jelly (RJ)/kg twice per week, respectively, over a 20-wk period. Semen quality and blood biochemical constituents were evaluated. RJ at any dose exhibited a significant increase (P<0.05) in rabbits’ sperm concentration, total sperm output, sperm motility, live sperm and normal sperm compared to the untreated controls. Plasma total protein, albumin, globulin, glucose and high density lipids (HDL) concentrations were significantly (P<0.05) boosted in the RJ groups compared to the controls. In contrast, RJ treatment resulted in a significant (P<0.05) reduction in plasma total lipids, triglycerides, cholesterol and low density lipids (LDL) concentrations. Treatment with RJ significantly boosted (P<0.05) testosterone concentration in the RJ groups to reach 110, 120 and 128%, respectively, of the control group. Improved kidney and liver functions were observed in the RJ bucks groups where plasma creatinine, urea concentrations, aspartate aminotransferase and alanine aminotransferase enzyme activities were significantly (P<0.05) decreased by RJ treatments. Treating bucks subjected to heat stress by different RJ doses increased (P<0.05) total antioxidant capacity to 106, 111 and 115% of basal, but significantly reduced (P<0.05) malondialdehyde and thiobarbituric acid reactive substances compared to the untreated. It was concluded that Chinese royal jelly supplementation for heat-stressed male rabbits can counteract summer infertility and improve their physiological status.

Protective effects of royal jelly on ofloxacin in male rat reproductive system are not well-known. This study was designed to determine the protective effects of royal jelly on the DNA integrity of sperms and early in vitro embryonic development in ofloxacin treated rats. In this study, 32 adult male rats were used and randomly allocated into 4 groups, each consisted of eight rats and dosed daily for 28 days by oral gavage: control—distilled water (216 mg/kg BW); ofloxacin group—ofloxacin (216 mg/kg BW); royal jelly group—royal jelly (100 mg/kg BW); royal jelly with ofloxacin group—216 mg/kg BW ofloxacin and 100 mg/kg BW royal jelly. The results showed that sperm viability, sperm population, sperm maturation, DNA maturity, hatching, IVF, GSH, and blastocyst significantly decreased (P < 0.05) in the ofloxacin group compared to the control. However, ofloxacin only caused significant increases (P < 0.05) in testis weight, immature sperm, and DNA damage of sperm compared to control group. In the ofloxacin with royal jelly group, there were not any important alterations detected and all sperm parameter returned to normal range. Royal jelly has protective effects on the DNA maturity of sperms and early in vitro embryonic development in ofloxacin treated rats.

The optimized capillary electrophoresis (CE) was applied to separate and detect the 10-hydroxy-2-decenoic acid (10-HAD) in royal jelly products. The method only requires that the sample solution need to be centrifuged and filtered before analyzed by the home-made capillary electrophoresis system. Firstly, 10-HDA was separated in a fused silica column with a diameter of 50 um using 20 mM Tris/Acetic buffer (pH 8.5) as a background electrolyte and a separation voltage of -17kV. Then, 10-HDA was detected by capacitively coupled contactless conductivity detection (C4D) with the migration time less than 8 minutes. Nine commercial products of royal jelly with Vietnamese and imported origin including pure royal jelly cream, lyophilized royal jelly (powder and gel) and honey with royal jelly were collected for analysis. The results of this study showed that content of 10-HDA were detected in the range of 0.5 mg/g to 23.1 mg/g. Using paired t test showed that the difference between results obtained from CE-C4D method and from HPLC method as a reference method was not statistically significant.&#x0D; Keywords: 10-HDA, royal jelly products, CE-C4D.&#x0D; References&#x0D; &#x0D; [1] M. Viuda Martos, Y. Ruiz Navajas, J. Fernández López, J.A. Pérez Álvarez, Functional properties of honey, propolis, and royal jelly”, Journal of food science 73 (2007) 117-124. https://doi:10.1111/j. 1750-3841.2008.00966.x.[2] M.F. Ramadan, A. Al-Ghamdi, Bioactive compounds and health-promoting properties of royal jelly: A review, Journal of Functional Foods 4 (2012) 39- 52. https://doi.org/10.1016/j.jff. 2011. 12.007.[3] Dimitrios Kanelis, Chrysoula Tananaki, Vasilis Liolios, Maria Dimou, Georgios Goras, Maria Anna Rodopoulou, Emmanuel Karazafris, and Andreas Thrasyvoulou, A suggestion for royal jelly specifcations, Archives of Industrial Hygiene and Toxicology 66 (2015) 275-284. Https://doi: 10.1515/aiht-2015-66-2651.[4] Brazil Ministério Da Agricultura e Do Abaste Imento. Secretaria De Defesa Agropecuária. Instrução normativa nº3, de 19 de janeiro de 2001. Regulamento técnico para fxação de identidade e qualidade de geléia real, Regulamento técnico para fxação de identidade e qualidade de geléia real liofilizada. Http:// www.engetecno.com.br/port/ legislacao/mel_geleia_real.htm.[5] Mahmut Genc¸ Abdurrahman Aslan, Determination of trans-10-hydroxy-2-decenoic acid content in pure royal jelly and royal jelly products by column liquid chromatography, Journal of Chromatography A. 839 (1999) 265 – 268. Http://doi: 10.1016/s0021-9673(99)00151-x.[6] F. Ferioli, E. Armaforte, M.F. Caboni, Comparison of the Lipid Content, Fatty Acid Profile and Sterol Composition in Local Italian and Commercial Royal Jelly Samples” Journal of the American Oil Chemists' Society, 91(2014) 875-884. Http://doi. org/10.1007/s11746-014-2446-x.[7] J. Kim, J. Lee, Quantitative analysis of trans-10-hydroxy-2-decenoic acid in royal jelly products purchased in USA by high performance liquid chromatography”, Journal of Apicultural Science, 54 (2010) 77-85. Https://pdfs.semanticscholar. org/e85f/dfc1823d778a40c ca2eefa4be8ba4b4d98 6c.pdf.[8] C.I. Pavel, L.A. Mărghitaş, D.S. Dezmirean, L.I. Tomoş, V. Bonta, A. Şapcaliu, A. Buttstedt, Comparison between local and commercial royal jelly-use of antioxidant activity and 10-hydroxy-2-decenoic acid as quality parameter”, Journal of Apicultural Research 53 (2014) 116-123. https:// doi.org/10.3896/IBRA.1.53.1.12.[9] Mahmut Genc¸ Abdurrahman Aslan, Determination of trans-10-hydroxy-2-decenoic acid content in pure royal jelly and royal jelly products by column liquid chromatography, Journal of Chromatography A, 839 (1999) 265 – 268. Https://doi:10.1016/s0021-9673(99)00151-x.[10] Jinhui Zhou, Xiaofeng Xue, Yi Li, Jinzhen Zhang and Jing Zhao, Optimized determination method for trans-10-hydroxy-2-decenoic acid content in royal jelly by high-performance liquid chromatography with an internal standard, Journal of AOAC International, 90(2007) 244-249. https://www.ncbi.nlm.nih.gov/pubmed/17373456.[11] Federico Ferioli, Gian Luigi Marcazzan, Maria Fiorenza Caboni, Determination of (E)-10-hydroxy-2-decenoic acid content in pure royal jelly: A comparison between a new CZE method and HPLC”, J. Sep. Sci .30 (2007) 1061-1069. Http://doi:10.1002/jssc.200600416.[12] Orlando Muñoz, Susana Decap, Francisco Ruiz, José Arbildua, Octavio Monasterio, Determination of 10- hydroxy-2-decenoic acid in royal jelly by capillary electrophoresis, J. Chil. Chem. Soc., 56 (2010) 738-740. http://dx.doi.org/10.4067/S0717 97072011000300004.

The genetic selection of a high royal jelly (RJ)-producing strain of bees (RJBs) from Italian bees (ITBs) has been conducted for nearly four decades in China since the 1980s. RJBs are the most important producers of RJ in the world and produce > 90% of the total output with an annual market value of > $40 million. With technological advancements in proteomics, the mechanism underpinning the high RJ production by RJBs has been explored to new depths in the last decades. Here, we give an overview that the mechanism driving the enhanced performance of RJ secretion by RJBs. First, the selection of RJBs, high-efficiency technology for RJ production, and advances in genetic characterization of RJBs are reviewed. Then, proteome and phosphoproteome characterization that decipher the augmented RJ production using honeybee organs and tissues are summarized. This may be potentially useful in gaining a complete mechanistic insight into the high performance of RJ yields in honeybees, and expands understanding of the honeybee biology.