MicroRNA profiling of royal jelly extracellular vesicles and their potential role in cell viability and reversing cell apoptosis.

MiRNAs are small non-coding RNA molecules that play important regulatory roles in diverse biological processes. Royal jelly, a milky-white substance produced by nurse honeybees (Apis mellifera), is the primary food of queen bees and plays a crucial role in their development. However, little is known about the microRNA (miRNAs) content of royal jelly and their potential functions. In this study, we isolated extracellular vesicles from the royal jelly of 36 samples through sequential centrifugation and targeted nanofiltration and performed high-throughput sequencing to identify and quantify the miRNA content of honeybee royal jelly extracellular vesicles (RJEVs). We found a total of 29 known mature miRNAs and 17 novel miRNAs. Through bioinformatic analysis, we identified several potential target genes of the miRNAs present in royal jelly, including those involved in developmental processes and cell differentiation. To investigate the potential roles of RJEVs in cell viability, RJEVs were supplemented to apoptotic porcine kidney fibroblasts induced by ethanol 6% exposure for 30 min. TUNEL assay showed a significant reduction in the apoptosis percentage after RJEV supplementation when compared with the non-supplemented control group. Moreover, the wound healing assay performed on the apoptotic cells showed a rapid healing capacity of RJEV-supplemented cells compared to the control group. We observed a significant reduction in the expression of the miRNA target genes such as FAM131B, ZEB1, COL5A1, TRIB2, YBX3, MAP2, CTNNA1, and ADAMTS9 suggesting that RJEVs may regulate the target gene expression associated with cellular motility and cell viability. Moreover, RJEVs reduced the expression of apoptotic genes (CASP3, TP53, BAX, and BAK), while significantly increasing the expression of anti-apoptotic genes (BCL2 and BCL-XL). Our findings provide the first comprehensive analysis of the miRNA content of RJEVs and suggest a potential role for these vesicles in the regulation of gene expression and cell survival as well as augmenting cell resurrection or anastasis.

ROCK Inhibitor (Y-27632) Abolishes the Negative Impacts of miR-155 in the Endometrium-Derived Extracellular Vesicles and Supports Embryo Attachment.

Extracellular vesicles (EVs) are nanosized vesicles that act as snapshots of cellular components and mediate cellular communications, but they may contain cargo contents with undesired effects. We developed a model to improve the effects of endometrium-derived EVs (Endo-EVs) on the porcine embryo attachment in feeder-free culture conditions. Endo-EVs cargo contents were analyzed using conventional and real-time PCR for micro-RNAs, messenger RNAs, and proteomics. Porcine embryos were generated by parthenogenetic electric activation in feeder-free culture conditions supplemented with or without Endo-EVs. The cellular uptake of Endo-EVs was confirmed using the lipophilic dye PKH26. Endo-EVs cargo contained miR-100, miR-132, and miR-155, together with the mRNAs of porcine endogenous retrovirus (PERV) and ß-catenin. Targeting PERV with CRISPR/Cas9 resulted in reduced expression of PERV mRNA transcripts and increased miR-155 in the Endo-EVs, and supplementing these in embryos reduced embryo attachment. Supplementing the medium containing Endo-EVs with miR-155 inhibitor significantly improved the embryo attachment with a few outgrowths, while supplementing with Rho-kinase inhibitor (RI, Y-27632) dramatically improved both embryo attachment and outgrowths. Moreover, the expression of miR-100, miR-132, and the mRNA transcripts of BCL2, zinc finger E-box-binding homeobox 1, ß-catenin, interferon-γ, protein tyrosine phosphatase non-receptor type 1, PERV, and cyclin-dependent kinase 2 were all increased in embryos supplemented with Endo-EVs + RI compared to those in the control group. Endo-EVs + RI reduced apoptosis and increased the expression of OCT4 and CDX2 and the cell number of embryonic outgrowths. We examined the individual and combined effects of RI compared to those of the miR-155 mimic and found that RI can alleviate the negative effects of the miR-155 mimic on embryo attachment and outgrowths. EVs can improve embryo attachment and the unwanted effects of the de trop cargo contents (miR-155) can be alleviated through anti-apoptotic molecules such as the ROCK inhibitor.

Vitamin C enhances porcine cloned embryo development and improves the derivation of embryonic stem-like cells.

Porcine cloning through somatic cell nuclear transfer (SCNT) has been widely used in biotechnology for generating animal disease models and genetically modified animals for xenotransplantation. Vitamin C is a multifunctional factor that reacts with several enzymes. In this study, we used porcine oocytes to investigate the effects of different concentrations of vitamin C on in vitro maturation (IVM), in vitro culture (IVC), and the derivation of nuclear transfer embryonic stem-like cells (NT-ESCs). We demonstrated that vitamin C promoted the cleavage and blastocyst rate of genetically modified cloned porcine embryos and improved the derivation of NT-ESCs. Vitamin C integrated into IVM and IVC enhanced cleavage and blastocyst formation (P < 0.05) in SCNT embryos. Glutathione level was increased, and reactive oxygen species levels were decreased (P < 0.05) due to vitamin C treatment. Vitamin C decreased the gene expression of apoptosis (BAX) and increased the expression of genes associated with nuclear reprogramming (NANOG, POU5F1, SOX2, c-Myc, Klf4, and TEAD4), antioxidation (SOD1), anti-apoptotic (Bcl2), and trophectoderm (CDX2). Moreover, vitamin C improved the attachment, derivation, and passaging of NT-ESCs, while the control group showed no outgrowths beyond the primary culture. In conclusion, supplementation of vitamin C at a dose of 50 µg/ml to the IVM and IVC culture media was appropriate to improve the outcomes of porcine IVM and IVC and for the derivation of NT-ESCs as a model to study the pre- and post-implantation embryonic development in cloned transgenic embryos. Therefore, we recommend the inclusion of vitamin C as a supplementary factor to IVM and IVC to improve porcine in vitro embryonic development.

Effects of vitamin C, vitamin E, selenium, zinc, or their nanoparticles on camel epididymal spermatozoa stored at 4 °C.

This study determined the effects of antioxidant supplementation and storage time at cool temperatures on the characteristics of epididymal camel spermatozoa. Camel testes were collected at the abattoir after animal slaughtering and kept at 4 °C during transportation and until processing (max 6 h). Spermatozoa were retrieved and diluted with SHOTOR extender, split in aliquots, supplemented with the following antioxidants: 200 µm/mL vitamin E, 1.0 g/L vitamin C, 1 µg/mL selenium nanoparticles, 50 µg/mL zinc nanoparticles, 2 µg/mL sodium selenite, and 100 µg/mL zinc sulfate, and stored at 4 °C for 2, 48, 96, and 144 h. The storage time significantly affected (P < 0.05) the sperms' motility and livability, the sperms' membrane integrity, and the percentages of cytoplasmic droplets as well as the percentage of morphologically normal spermatozoa. Epididymal sperm characteristics (progressive motility, livability, membrane integrity, and abnormalities) were significantly improved (P < 0.05) when the spermatozoa were diluted with antioxidants as compared with the control group, and the best additives were identified as nano-selenium, sodium selenite, nano-zinc, and zinc sulfate. In conclusion, adding nano-sized minerals or inorganic trace elements and vitamins maintained the progressive motility, livability, and membrane integrity, and decreased abnormalities and cytoplasmic droplet percentages of epididymal camel spermatozoa stored at 4 °C up to 144 h.

Combined Supplementation of Nano-Zinc Oxide and Thyme Oil Improves the Nutrient Digestibility and Reproductive Fertility in the Male Californian Rabbits.

The present study aimed to determine the effects of zinc oxide nanoparticles (ZnO-NPs), thyme oil (THO), or their combination on the nutrient digestibility coefficients, reproductive parameters, and some blood metabolites of male Californian rabbits. One hundred rabbits, 29-weeks of age (initial body weight 3.48 ± 0.08 kg) were randomly distributed into four groups, 25 rabbits each. Treatment groups were fed a control diet, a control diet supplemented with ZnO-NPs (100 mg/kg), THO (500 mg/kg), or combination of ZnO-NPs (100 mg/kg) and THO (500 mg/kg). The feeding trial lasted for 35 days. Results showed improvements in dry matter, crude protein, ether extract, and crude fiber in ZnO-NPs, THO, and their combination treated groups compared to those of control. Furthermore, semen volume, sperm motility, vitality, and morphology were significantly improved (p < 0.01) in ZnO-NPs and THO groups rather than the control. Both ZnO-NPs and THO, as either individual or combined treatments significantly improved the serum alanine amino-transferase (ALT), aspartate amino-transferase (AST), urea, and creatinine compared to the control. Moreover, serum concentrations of testosterone were significantly increased in rabbits supplemented with ZnO-NPs, THO, or their combination compared to those of control (p < 0.05). In conclusion, ZnO-NPs, THO, or their combination improved the digestibility of nutrients, liver/ kidney functions, semen characteristics, and testosterone concentration in male rabbits.

Effects of mint, thyme, and curcumin extract nanoformulations on the sperm quality, apoptosis, chromatin decondensation, enzyme activity, and oxidative status of cryopreserved goat semen.

Goat semen cryopreservation is a challenging process as it results in reduced motility, vitality, and fertility of spermatozoa after freezing. In this study, we evaluated the effects of different herbal extract nanoformulations (NFs) [mint (MENFs), thyme (TENFs), and curcumin (CENFs)], supplemented at either 50 or 100 µg into Tris-extender on the cryopreserved goat semen quality. The hydrothermal squeezing method was used for the preparation of the NFs extracts. The morphological evaluation of the NFs extracts was conducted by transmission electron microscopy. All NFs supplements improved (p < 0.05) the progressive motility, vitality, and plasma membrane integrity of sperm compared with the control extender after equilibration (5 °C for 2 h) and thawing (37 °C for 30 s), but had no effect on sperm abnormality and acrosome integrity. All NFs supplements decreased (p < 0.05) the apoptosis, malondialdehyde level, and chromatin decondensation of sperm cells, while increased (p < 0.05) the total antioxidant capacity and catalase activity in the frozen/thawed extender. Particularly, CENFs at a level of 100 µg showed improvement of sperm parameters and antioxidant status during cryopreservation of goat semen more than TENFs and MENFs. The CENFs improved the quality of goat spermatozoa in post-thawed semen in terms of preventing cryodamage and promoting the cryotolerance of spermatozoa when compared with TENFs and MENFs. Therefore, supplementation of Tris-extender with CENFs could enhance goat semen processing during cryopreservation.

Comparison between the Effects of Adding Vitamins, Trace Elements, and Nanoparticles to SHOTOR Extender on the Cryopreservation of Dromedary Camel Epididymal Spermatozoa.

There are several obstacles in camel semen cryopreservation; such as increasing semen viscosity and the reduction in motile spermatozoa after ejaculation. Epididymal spermatozoa offer an efficient alternative to overcome these problems and are well-suited for artificial insemination in camels. In the current study, we compared the effects of supplementation with vitamin C, E, inorganic trace elements of selenium (Na2SeO3) and zinc (ZnSO4), and zinc and selenium nanoparticles (ZnONPs and SeNPs, respectively) on the cryopreservation of dromedary camel epididymal spermatozoa. When the SHOTOR extender was supplemented with ZnONPs and SeNPs; the sperm showed increased progressive motility; vitality; and membrane integrity after cooling at 5 °C for 2 h; when compared to the control and vitamin-supplemented groups. Moreover, the ZnONPs and SeNPs supplementation improved the progressive motility, vitality, sperm membrane integrity, ultrastructural morphology, and decreased apoptosis when frozen and thawed. SeNPs significantly increased reduced glutathione (GSH), superoxide dismutase (SOD), and decreased lipid peroxide malondialdehyde (MDA) levels. The advantageous effects of the trace elements were potentiated by reduction into a nano-sized particle, which could increase bioavailability and reduce the undesired liberation of toxic concentrations. We recommend the inclusion of SeNPs or ZnONPs to SHOTOR extenders to improve the cryotolerance of camel epididymal spermatozoa.

Effects of supplementing broiler diets with coriander seed powder on growth performance, blood haematology, ileum microflora and economic efficiency.

A total of 480 one-day-old Arbor Acres broiler chicks were randomly assigned to four dietary groups, each including six replicates (n = 20/replicate). Broilers in the first group (G1) were fed a basal diet without any additives (control). Broilers in groups 2, 3 and 4 (G2, G3 and G4) were fed a basal diet supplemented with 0.1%, 0.2% and 0.4% coriander (Coriandrum sativum L.) seed powder (CSP) respectively. Feeding trials lasted for 42 days, and after that growth, carcass traits, haematological parameters, gut microbiota and economic efficiency (EE) were evaluated. Final body weight (FBW), total weight gain (TWG), total feed intake (TFI) and red blood cell (RBC) counts of broilers in the G4 and G3 treatment groups were significantly higher (p < .05) compared with broilers in the G1 treatment group. The feed conversion ratio (FCR) was better (p < .05) in the G4 treatment group (1.72) than in the G1 treatment group (1.84). Broilers in CSP treatment groups had significantly higher haemoglobin (Hb) concentrations, packed cell volume (PCV) and platelet counts, and had lower (p < .05) Escherichia coli and Clostridium perfringens counts compared with broilers in the G1 treatment group. Dressing, liver, pancreas, bursa and thymus percentages were higher (p < .05) in broilers in the G4 (70.30, 3.18, 0.31, 0.10 and 0.32% respectively) treatment group, compared with broilers in the control (G1) group (66.57, 2.37, 0.23, 0.04 and 0.21% respectively). Broilers in treatment groups G4 and G3 had lower percentages of abdominal fat and lower total bacterial counts (p < .05) than broilers in treatment groups G2 and G1. The highest economic efficiency (EE) was found in treatment group G4, and EE in this group was 13.06% greater than in the control (G1) group.

The Usefulness of Retinoic Acid Supplementation during In Vitro Oocyte Maturation for the In Vitro Embryo Production of Livestock: A Review.

Retinoic acid (RA) is an indigenous metabolite and descriptive physiologically functioning constituent of vitamin A. Retinoids were documented as vital regulators for cell development and distinction, embryonic growth, and reproductive function in both male and female livestock. Previously, RA has been shown to have several positive impacts in vivo and in vitro and critically control many reproductive events, such as oocyte development, follicular growth, and early embryonic growth. In addition, RA manages apoptotic signaling and oxidative damages in cells. Recently, RA has been used widely in assisted reproductive technology fields, especially during in vitro embryo development in various mammalian species, including buffaloes, bovine, goats, sheep, pigs, and rabbits. However, the optimum concentration of RA greatly differs based on the condition of maturation media and species. Based on the obtained findings, it was generally accepted that RA enhances nuclear oocyte maturation, cleavage and maturation rates, blastocyst formation, and embryo development. As such, it possesses antioxidant properties against reactive oxygen species (ROS) and an anti-apoptotic effect through enhancing the transcription of some related genes such as superoxide dismutase, prostaglandin synthase, glutathione peroxidase, peroxiredoxins, and heme oxygenase. Therefore, the current review concludes that an addition of RA (up to 50 nM) has the potential to improve the oocyte maturation media of various species of livestock due to its antioxidant activity.

Endosulfan toxicity in Nile tilapia (Oreochromis niloticus) and the use of lycopene as an ameliorative agent.

OBJECTIVES: Endosulfan is a broad-spectrum organochlorine insecticide that has been commercially in use for decades to control insect pests and has been found to pollute the aquatic environment. The current study was carried out to investigate the toxic effects of endosulfan, an organochlorine pesticide, on Nile tilapia (Oreochromis niloticus), a freshwater fish, and the alleviating effects of lycopene on the induced toxicity. METHODS: Four treatment groups of fish were investigated (3 replicates of 15 fish for each group): (1) a control group, (2) a group exposed to endosulfan, (3) a group that was fed on a basal diet supplemented with lycopene, and (4) a group that was fed on a basal diet supplemented with lycopene and exposed to endosulfan. The experiment was carried out over a 4-week period. RESULTS: Endosulfan negatively affected liver function, including liver enzymes and plasma proteins. Endosulfan affected blood parameters of fish and reduced the counts of red blood cells (RBCs) and white blood cells (WBCs), as well as affected immunological parameters. Endosulfan caused oxidative stress, as it decreased the values of antioxidants catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPX), and glutathione (GSH), and increased the level of lipid peroxide malondialdehyde (MDA). Additionally, endosulfan increased cytochrome P450 (CYP450) levels, while it decreased glutathione S-transferase (GST) mRNA transcript levels and distorted the normal histological structure of the liver, gills, and spleen of affected fish. Conversely, lycopene partially restored the aforementioned parameters when administered concomitantly with endosulfan. CONCLUSION: The results showed the beneficial effects of supplementing fish diets with lycopene as a natural antioxidant for ameliorating the toxicity caused by endosulfan.

Effects of all-trans retinoic acid on the in vitro maturation of camel (Camelus dromedarius) cumulus-oocyte complexes.

All-trans retinoic acid (RA) is a metabolite of vitamin A and has pleiotropic actions on many different biological processes, including cell growth and differentiation, and is involved in different aspects of fertility and developmental biology. In the current study, we investigated the effects of RA on camel (Camelus dromedarius) cumulus-oocyte complex in vitro maturation (IVM). IVM medium was supplemented with 0, 10, 20, and 40 µM RA. Application of 20 µM RA significantly reduced the proportion of degenerated oocytes and significantly improved oocyte meiosis and first polar body extrusion compared to the control and other experimental groups. Retinoic acid significantly reduced the mRNA transcript levels of apoptosis-related genes, including BAX and P53, and reduced the BAX/BCL2 ratio. In addition, RA significantly reduced the expression of the Transforming growth factor beta (TGFß) pathway-related transcripts associated with the actin cytoskeleton, ACTA2 and TAGLN; however, RA increased TGFß expression in cumulus cells. The small molecule SB-431542 inhibits the TGFß pathway by inhibiting the activity of activin receptor-like kinases (ALK-4, ALK-5, and ALK-7); however, combined supplementation with RA during IVM compensated for the inhibitory effect of SB-431542 on cumulus expansion, oocyte meiosis I, and first polar body extrusion in activated oocytes. The current study shows the beneficial effects of RA on camel oocyte IVM and provides a model to study the multifunctional mechanisms involved in cumulus expansion and oocyte meiosis, particularly those involved in the TGFß pathway.

Impact of Rosmarinus officinalis cold-pressed oil on health, growth performance, intestinal bacterial populations, and immunocompetence of Japanese quail.

This study was performed to study the impacts of rosemary cold-pressed oil (RCPO) for biostimulating health, growth performance, and intestinal bacterial populations of Japanese quail. The study included 300 growing 1-wk-old Japanese quails. Birds were divided into 3 groups in a complete randomized design experiment that involved 3 levels of RCPO (0, 1.00 and 2.00 mL/kg diet). Results revealed that the addition of rosemary oil numerically increased values of body weight and body weight gain when compared with the control group, particularly the highest level of RCPO (2.00 mL/kg diet). Birds fed diets supplemented with rosemary oil consumed more feed (P ≤ 0.01) compared with those fed the control diet. Feed conversion ratio tended to be improved in rosemary oil groups during the period 3 to 6 wk of age (P = 0.013). The highest level of rosemary oil (2.00 mL/kg diet) had the best impact on all carcass traits studied. RCPO supplementation showed an increase in serum total protein, metabolic hormones levels, while it reduced serum cholesterol and low-density lipoprotein cholesterol, 8-hydroxy-2'-deoxyguanosine, and protein carbonyl levels. Moreover, RCPO increased antioxidative enzymes, and reduced the lipid peroxidation in quail liver. The supplementation of 2 mL/RCPO kg diet showed significant reduction in populations of total cultural bacterial count, coliforms, Escherichia coli, and Salmonella spp. in the ileum when compared to the control. The current results showed that RCPO supplementation to Japanese quails diet could enhance the growth performance and reduce the intestinal pathogenic bacteria. Therefore, RCPO can be a beneficial antimicrobial and growth-promoting feed supplement for the Japanese quail.

Efficacy of Montanide (IMS 3015) as an adjuvant for an inactivated Rift Valley fever (RVF) vaccine in sheep.

This study was conducted to evaluate an adjuvant, Montanide (IMS 3015), in improving the quality of Rift Valley fever (RVF) vaccine relative to the traditional adjuvant, aluminum hydroxide gel. Vaccinated sheep were evaluated using biochemical analysis, kidney function tests, liver function tests, and immunological tests. Sheep vaccinated with Montanide (IMS 3015) adjuvant showed significantly higher total protein, total globulin, and gamma globulin concentrations from the second week until the fifth month than the controls. Conversely, albumin concentration and the A/G ratio significantly decreased during this period. Kidney function and liver function tests revealed no differences among any of the groups. There was a significant increase in lymphocyte proportion and a decrease in neutrophil proportion in sheep vaccinated with the Montanide (IMS 3015) adjuvant. Lymphocyte cell proliferation was significantly different in sheep vaccinated with the Montanide (IMS 3015) adjuvant from that in controls. Neutralizing indices were significantly higher in sheep vaccinated with the Montanide (IMS 3015) adjuvant than in controls. The current study showed that sheep vaccinated with inactivated RVF virus with Montanide (IMS 3015) as an adjuvant were protected and no pathological symptoms or biochemical changes were detected. Moreover, the vaccine induced rapid onset of immunological responses with long durations unlike inactivated RVF vaccine with aluminum hydroxide gel.

Effects of clove (Syzygium aromaticum) oil on quail growth, carcass traits, blood components, meat quality, and intestinal microbiota.

To provide a safe growth promoter, the present study has investigated the effects of different levels of cold-pressed clove oil (CCPO) on growth performance, carcass traits, blood biochemistry, and intestinal microbial population of growing Japanese quails. A total of 300 quails (1-wk old) were randomly divided into 3 treatment groups: control basal diet, basal diet +0.75 mL oil/kg diet, and basal diet +1.5 mL oil/kg diet. Quails fed with 1.5 mL clove oil/kg diet showed a 3.43% improvement in live body weight vs. control group. Similar trend was observed for daily body weight gain. Feed intake gradually increased (P < 0.01) with an increase in clove oil level. The best feed conversion rate was reported for the control group, followed by the group treated with 1.5 mL CCPO/kg diet during the whole period (1 to 6 wk of age). Total globulin differed significantly in 1.5 mL CCPO/kg diet group. Antioxidant enzyme activities, lipid profile, and reduced glutathione concentrations significantly improved in a dose-dependent manner. Blood urea nitrogen, creatinine, malondialdehyde, 8-hydroxy-2΄-deoxyguanosine, and protein carbonyl levels significantly decreased in quails supplemented with 1.5 mL CCPO/kg diet vs. control group. Serum levels of insulin-like growth factor-1, insulin, growth hormone, and thyroxine significantly increased in quails supplemented with 1.5 mL CCPO/kg diet vs. control group. The intestinal bacterial population, coliforms, Escherichia coli, and Salmonella spp. in the ileal content were lower (P < 0.05) in groups treated with oil (1.5 mL/kg) vs. control group. Thus, dietary supplementation with antimicrobial CCPO (1.5 mL/kg diet) could enhance growth performance, improve health status, and reduce intestinal pathogens in Japanese quails.

Ameliorative effect of ginseng extract on phthalate and bisphenol A reprotoxicity during pregnancy in rats.

Phthalates (such as DEHP) and bisphenol A (BPA) are widely used chemicals in plastics manufacturing and exert public health concerns as endocrine disrupters. This study was designed to investigate the deleterious effect of DEHP and BPA on endocrine profile of pregnant female rats and the combined treatment with ginseng extract (Panax ginseng). Seventy-two pregnant rats were divided into six groups (control, ginseng, DEHP, BPA, Gin + DEHP, and Gin + BPA), 12 females per each group. The drugs were supplemented from pregnancy day 0 until day 20. Determination of serum sex hormones (testosterone, progesterone, and estradiol) were determined on days 4, 10, and 20 of pregnancy. mRNA transcripts of STAR, HSD17B3, CYP17, AKT1, and PTEN were relatively quantified against ACTB in the ovary and placenta of days 10 and 20 pregnant females by relative quantitative polymerase-chain reaction (RQ-PCR). DEHP and BPA significantly decreased the endocrine profile of testosterone, progesterone, and estradiol of days 10 and 20 of pregnant females. Combined administration of these chemicals along with ginseng extracts has returned the hormones to normal levels when compared with the control group. The ovarian and placental CYP17 and HSD17B3 mRNA transcripts showed variable expression pattern in both tissues and they were significantly affected by DEHP and BPA administration, concomitantly correlating to STAR, AKT1, PTEN, progesterone, and testosterone levels on pregnancy days 10 and 20. The results confirm the reprotoxicity of DEHP and BPA as endocrine disruptors and indicate that ginseng could be used to alleviate the toxic effects of these chemicals.

Impacts of supplementing growing rabbit diets with whey powder and citric acid on growth performance, nutrient digestibility, meat and bone analysis, and gut health.

The present study evaluated the impact of supplementing the rabbit diet with graded levels of whey powder and citric acid. The dietary treatments were as follows: T1, control diet (basal diet); T2, basal diet + 10 g/kg citric acid; T3, T2 + whey powder (7.5 g/kg); T4, T2 + whey powder (15 g/kg); and T5, T2 + whey powder (22.5 g/kg). Results, the T5 diet resulted in the best (P < 0.05) final body weight, body weight gain, feed conversion ratio, protein efficiency, relative growth rate, and dressed weight. The best (P < 0.05) digestion coefficients were associated with the T4 and T5 diets. Rabbits fed diets supplemented with citric acid alone or with addition of graded levels of whey powder showed significantly lower (P < 0.05) intestinal pH than those fed the T1 diet. The T4 and T5 diets resulted in greater CP and ash in the thigh muscle compared with the T1 and T2 diets. Calcium content in the femur bone was higher (P < 0.05) in the T5 group followed by T4 and T3. The wall of different parts of the small intestine improved in the T4 and T5 groups, showing the greatest increase in the small intestinal villi, intestinal glands, and amount of goblet cells. In conclusion, addition of whey powder (1.5, and 2.25%) increased the growth performance, nutrient digestibility and crude protein content of the thigh muscle, and improved the gut health of growing rabbits and the best level was 2.25% whey powder. Citric acid addition had no positive effect on growth performance, nutrient digestibility, crude protein content of the thigh muscle, and the gut health.

Effect of Kisspeptin on the Developmental Competence and Early Transcript Expression in Porcine Oocytes Parthenogenetically Activated with Different Methods.

Recent studies showed the modulatory effect of kisspeptin (KP) on calcium waves through the cell membrane and inside the cell. Spermatozoon can induce similar ooplasmic calcium oscillations at fertilization to trigger meiosis II. Here, we evaluated the effect of KP supplementation with 6-dimethylaminopurine (6-DMAP) for 4 h on embryonic development after oocyte activation with single electric pulse, 5 µM ionomycin, or 8% ethanol. Compared to control nonsupplemented groups, KP significantly improved embryo developmental competence electric- and ethanol-activated oocytes in terms of cleavage (75.3% and 58.6% versus 64% and 48%, respectively, p < 0.05) and blastocyst development (31.3% and 10% versus 19.3% and 4%, respectively, p < 0.05). MOS expression was increased in electrically activated oocytes in presence of KP while it significantly reduced CCNB1 expression. In ionomycin treated group, both MOS and CCNB1 showed significant increase with no difference between KP and control groups. In ethanol-treated group, KP significantly reduced CCNB1 but no effect was observed on MOS expression. The early alterations in MOS and CCNB1 mRNA transcripts caused by KP may explain the significant differences in the developmental competence between the experimental groups. Kisspeptin supplementation may be adopted in protocols for porcine oocyte activation through electric current and ethanol to improve embryonic developmental competence.

Improving growth performance and health status of meat-type quail by supplementing the diet with black cumin cold-pressed oil as a natural alternative for antibiotics.

Using antibiotics in poultry diets as growth promoters was reported to have harmful effects on consumers, so the current study was done to monitor the impact of dietary supplementation of antimicrobial black cumin oil (BCO) on carcass traits, growth performance, biochemical components, and ileal microbial populations of growing Japanese quails. Three hundred growing Japanese quails were used with three different treatments (0, 0.50, and 1.0 g BCO/g diet). Birds fed diet supplemented with 0.5 g BCO/kg diet showed significant increase in body weight comparing with the control and other treatment group. The daily feed intake and feed conversion ratio were significantly increased side by side with increasing BCO level in the diet. The majority of carcass characteristics were maximized by supplementing the quail diet with 0.5 g BCO/kg. Moreover, liver functions, anti-oxidative capacity, lipid profile and anabolic hormones showed significant improvement in BCO-treated diets in a dose-dependent manner. The BCO showed highest antibacterial effect against Escherichia coli and Salmonella enterica. The ileal bacterial populations, i.e., total bacterial count (TBC), coliform, Salmonella species, and Escherichia coli were decreased in birds supplemented with BCO 0.5 and 1.0 BCO g/kg compared with the control diet. Based on the aforementioned results, conclusion could be drawn that supplementing quail with BCO in their diet could improve productive performance traits and enhance health aspect of the birds.

Alleviating the environmental heat burden on laying hens by feeding on diets enriched with certain antioxidants (vitamin E and selenium) individually or combined.

The present study was designed to alleviate the negative biohazards of high ambient temperature on the productive performance and physiological status of laying hens. A total of 135 Bovans laying hens were distributed into nine groups in a 3 × 3 factorial design experiment. Basal diet was supplemented with vitamin E at levels of 0, 250, and 500 mg /kg diet. Within each dietary vitamin E level, each diet was supplemented with sodium selenite as a source of selenium (Se) to supply 0, 0.25, and 0.50 mg Se/kg diet. Results showed that supplementing layer's diet with 500 mg vitamin E/kg was accompanied with the lowest feed consumption (FC) and feed conversion ratio (FCR). The interaction among vitamin E and Se levels exerted significant effects only on FC and FCR. Insignificant differences were observed in egg quality criteria due to the treatments studied. Increasing vitamin E level was associated with a gradual decrease in basophil count and an increase in monocytes. A gradual decrease in the count of each of heterophils, monocytes, and eosinophils was observed with the elevation in the dietary Se level. The combination among vitamin E and Se levels produced a significant effect on all hematological parameters studied. As vitamin E increased, a marked decrease in serum AST and a gradual increase in total lipids, total cholesterol, and calcium were observed. As the level of dietary Se increased, serum total protein, albumin, T4, total cholesterol, and total lipids increased. No significant impacts were detected for the interaction among vitamin E and Se levels on any of blood constituents determined except serum globulin, ALT, and calcium. In conclusion, the combination between vitamin E and Se showed a good ability to alleviate the harmful impacts of heat stress and produced the highest productive performance when compared with the other groups, which exhibit the synergistic effect between the two antioxidants.