Complementary assessment of nano-packaged garlic properties by electronic nose.

It is crucial to initiate appropriate storage conditions for garlic depending on its properties. Fungal contamination can reduce the quality of garlic through changes in its properties which result in its aroma alteration. This study aimed to evaluate the effects of treatments such as fungal infection (FI), material of packaging (MP), and storage duration (SD) on various characteristics of garlic. An electronic nose was used complementarily to trace the aroma changes as a non-destructive indicator. The Fusarium oxysporum (FS), Alternaria embellisia (AL), and Botrytis allii (BT) fungi were adopted for inoculation. The low-density polyethylene (LDPE) and silicone nano-emulsions (SNE) were used for packing samples. The data were analyzed by diverse approaches such as ANOVA, PLS, PCA, LDA, and BPNN. The results revealed that the evaluated properties changed during the storage. The implementation of treatments altered the intensity of these changes. The highest values of weight loss (21.14%), color changes (50.21), and acidity (7.48) were observed in the FS-infected samples kept in LDPE for 28 days. The accuracy of PCA, LDA, and BPNN in the multivariate analysis of aroma had an increasing-decreasing trend. The best accuracy of PCA in categorizing the FI and MP treatments achieved in the twelfth day of storage (96%). The optimal accuracy of classifications based on FI and MP treatments was obtained at d#12 (100%) and d#24 (100%), respectively. The PLS exposed that the aroma changes in garlic had a high correlation with the changes of studied properties (R 2 ≥ .7), except for the mechanical properties.

Vitamin C Synergistically Enhances Protective Effects of Vitamin E Against Preantral Follicle Degeneration of Ovine Vitrified/Warmed Ovarian Tissue.

This study aimed to evaluate whether the addition of vitamins E and C as two conventional antioxidants improves the cryotolerance of preantral follicles enclosed in ovine ovarian tissue slices. For this purpose, ovarian slices were obtained from abattoired juvenile lambs and randomly distributed to the following groups: fresh, toxicity, vitrified (control), and three treatment groups in two experiments. Vitamin E, vitamin C, or vitamin E + C was added to the vitrification media alone in the first experiment and added to all vitrification, warming, and culture media in the second experiment. Finally, the treated tissues were cultured in vitro for 12 hours. The histological analysis showed that single or combined use of vitamins E and C increases intact preantral follicles in comparison to the control in two experiments (p < 0.05), and simultaneous use of vitamins E and C had a synergistic effect on increasing the percentage of normal preantral follicles in experiment 2 (p < 0.05). Due to the better results in Experiment 2, stromal cell density, antioxidant activity, and molecular evaluation were followed only in this experiment. The vitamin E + C group had higher stromal cell density compared with control group (p < 0.05). Vitamin E strengthened antioxidant capacity compared with the control and vitamin C groups (p < 0.05). This effect was exacerbated when used in combination with vitamin C (p < 0.05). The expression of all evaluated genes (BMP4, BMP15, GDF9, and KITLG) was significantly increased in ovarian tissue treated with vitamin E + C compared with the control group (p < 0.05). This increase was also observed in BMP4, GDF9, and KITLG genes compared with the vitamin C group (p < 0.05). In conclusion, this study revealed the positive effects of vitamins E and C on preantral follicle viability and to some extent a synergistic action of vitamin C on the protective effects of vitamin E against preantral follicle degeneration and increasing antioxidant capacity and development of preantral follicles after ovine ovarian tissue vitrification.

Potential use of cold plasma treatment for disinfection and quality preservation of grape inoculated with Botrytis cinerea.

Gray mold caused by Botrytis cinerea is a serious disease of grape (Vitis vinifera) during storage. The aim of this study is to evaluate the effect of atmosphere cold plasma (a novel and nonthermal technology) on inactivation of B. cinerea and preservation of chemical, physical, and mechanical characteristics of grape inoculated with B. cinerea. Herein, different time of cold plasma (0, 10, 20, and 40 s) was firstly considered to be the main factors, besides different storage time (1, 2, 3, 4, and 5 weeks) at 4°C. According to the results, plasma treatment exhibited inhibitory effect on gray mold percentage and microbial load of B. cinerea (log CFU g-1) during postharvest storage. So, in the last week, the gray mold percentage and microbial load in the control were 100% and 3.6 log CFU g-1, and in 40-s plasma were 4.5% and 2.53 log CFU g-1, respectively. Although the minimum infection and microbial load were observed in 40-s plasma, better postharvest quality preservation was observed in short-time cold plasma treatment (≤20 s). Forty-second plasma caused fruit tissue destruction and negatively decreased mechanical indices (Emod: 0.0028, Fmax = 1.78, and W = 3.18) and weight loss (91.9) in comparison with ≤20-s plasma, in which mechanical indices (Emod =0.0077, Fmax = 3.6, and W = 10.06) and weight loss (1/1) were higher. The long-time cold plasma treatment (40 s) had also maximum effects on color changes (10) and surface temperature (2.8°C). Although the highest TSS and TA were observed in 40-s Plasma, but different time of plasma treatments had no effect on pH. Altogether, these results indicate that the short-time cold plasma treatment can inactivate B. cinerea on grape berries and preserve crop quality properties.

Polyvinyl alcohol addition to freezing extender can improve the post-thaw quality, longevity and in vitro fertility of ram epididymal spermatozoa.

Recovering and cryopreserving epididymal spermatozoa are suitable methods for preserving the genetic potential of livestock and endangered species. Regarding encouraging reports on the use of polyvinyl alcohol (PVA) in cryopreserving various cell types, we conducted this study to examine the impact of PVA on the post-thaw quality, longevity, and in vitro fertility of ram epididymal sperm. In the first experiment, ram epididymal spermatozoa were frozen in extenders containing 6 % glycerol and 0, 0.5, 1, 2, 5, 10, or 15 mg/ml of PVA. Polyvinyl alcohol at concentrations of 0.5, 1, and 2 mg/ml improved the motility and functional membrane integrity (FMI) of the sperm compared with the control group (P < 0.05). In the second experiment, we investigated whether PVA could partially substitute glycerol in the freezing extender. PVA was added at 0, 0.5, 1, and 2 mg/ml to the extenders containing 1 % or 2 % glycerol. After thawing, the sperm motility parameters of the group containing 1 mg/ml PVA and 2 % glycerol were significantly higher than those of the un-supplemented groups (P < 0.05). In the third experiment, the effect of PVA on the post-thaw sperm longevity were examined. Sperm were frozen in 3 extenders: one containing 6 % glycerol and 1 mg/ml PVA (Gly6P1), another containing 2 % glycerol and 1 mg/ml PVA (Gly2P1), and a control extender with 6 % glycerol. After thawing, the quality of the sperm was evaluated. Sperm were then diluted in human tubal fluid (HTF) and incubated at 37 °C for 3 h. Afterwards, the quality of the sperm was evaluated once more. The presence of PVA in the freezing extender improved motility parameters and FMI. Additionally, PVA-containing groups had lower proportions of capacitated and acrosome reacted sperm compared with the control group (P < 0.05). The Gly6P1 group performed better than the other two groups (P < 0.05). In the fourth experiment, sperm from the Gly6P1 and Control groups were used in the IVF process immediately after thawing (T0) and after a 3-h incubation at 37 °C in HTF (T3). Cleavage, blastocyst and hatching rates in both groups were similar at T0, but they were lower in the Control group at T3 (P < 0.05). In conclusion, PVA as an additive to the freezing extender significantly improves post-thaw motility, viability, acrosome integrity, longevity, and fertile lifespan of ram epididymal spermatozoa.

Nanomagnetic Cyclodextrin decorated with ionic liquid as green and reversible Demulsifier for breaking of crude oil emulsions.

Application of the chemical demulsifiers is the best choice for breaking the water in crude oil (W/O) emulsions in the petroleum industry. Here, novel, environmentally friendly, efficient, and easily reusable Fe3O4 nanomagnetic compounds based on imidazolium-decorated cyclodextrin were successfully synthesized and applied to demulsify the water in crude oil (W/O) emulsions. At first, Fe3O4 nanoparticles were decorated with ß-cyclodextrin (ß-CD) to prepare Fe3O4@ß-CD@IL magnetic nanoparticles. Then, imidazole (Im) was separately reacted with 1-bromohexane and 1-bromodecane to prepare [Im-C6][Br] and [Im-C10][Br] ionic liquids, respectively. The prepared imidazolium-based ionic liquids were reacted with N-propyltriethoxysilane to synthesize [ImSi-C6][Br] and [ImSi-C10][Br]. Finally, [ImSi-Cn][Br] was immobilized on Fe3O4@ß-CD to obtain nanomagnetic demulsifiers. Structure of the synthesized compounds was confirmed using different methods such as FT-IR, NMR, and elemental analysis. TGA, VSM, and FESEM methods were used to investigate the thermal stability, magnetic properties, and the morphology, respectively. Fe3O4@ßCD and Fe3O4@ßCD@[ImSi-C10][Br] nanoparticles respectively showed the particle size in the range of 40-70 nm and 50-80 nm. After grafting the imidazolium-based ionic liquid on the surface of support, the magnetization number reduced from 25.6 emu/g for Fe3O4@ß-CD to 24.9 emu/g for Fe3O4@ß-CD@[ImSi-C10][Br]. Synthesized material employed to break the (10:90 and 30:70 Vol%) W/O emulsions at the concentration range of 1000-5000 ppm. The maximum demulsification efficiency (DE%) of 92 % was obtained using a Fe3O4@ß-CD@[ImSi-C10][Br] at 5000 ppm for (30:70 Vol%) W/O emulsion within 24 h. Interfacial tension (IFT) values decreased with increasing the DE%. The Fe3O4@ßCD@[ImSi-C10][Br] demulsifier was reused five times with acceptable yields. The cooperation of imidazolium and ß-CD in the green nanomagnetic demulsifiers led to the efficient demulsification of the W/O emulsions. The preparation of different ionic liquids or changing the counter anions are our potential future directions for this research. Demulsification at high demulsifier concentration can be considered a limitation of the nanomagnetic cyclodextrin decorated with ionic liquid. But due to the low amount of ionic liquid immobilized in the synthesized demulsifier, the cost of the final demulsifier is lower that other demulsifiers with full ionic liquid backbones, which increases its potential for industrial applications.

Effect of basil seed and xanthan gum on physicochemical, textural, and sensory characteristics of low-fat cream cheese.

This study aims to produce fat-reduced cream cheese using the different levels (0.25%-0.5%) of basil seed and xanthan gum by a RSM method. The basil seed, xanthan gum, and fat levels did not significantly influence the cream cheese's pH and acidity. With the fat reduction, textural properties were lost; for example, hardness, gumminess, and adhesiveness increased, and cohesiveness decreased. In addition, low-fat cream cheese's sensory score (taste, mouthfeel, and overall acceptance score) was lower. However, adding basil seed and xanthan gum could improve water holding capacity (WHC), hardness, gumminess, cohesiveness, adhesiveness and scores of mouthfeel, and overall acceptance. Basil seed gum had a better impact than xanthan on fat-reduced cream cheese properties among the two gums. In general, results showed that adding 0.5% basil and 0.5% xanthan into cream cheese could manufacture a product with a reduced-fat level (19.04%). At the same time, its physicochemical, sensory, and textural attributes were similar to cream cheese with high fat (24%). In addition, the price of the obtained product was lower.

The potential protective effect of melatonin and N-acetylcysteine alone and in combination on opioid-induced testicular dysfunction and degeneration in rat.

Methadone (Met) is the most common treatment for opioid addiction. Although Met is effective for treatment of opioid dependence, sexual dysfunctions and infertility have been reported as a major problem in patients under Met treatment. The present study aimed to evaluate the effect of melatonin and N-acetylcysteine (N) on morphine and Met-induced oxidative stress, apoptosis, suppression of blood sexual hormones, impairment in sperm parameters, and sexual dysfunction. Adult male Wistar rats (n = 66) were randomly divided into 11 equal groups (n = 6) as follows: control, sham, morphine, Met, Met+N, Met+ melatonin, Met+melatonin+N, morphine+ Met, morphine+Met+ melatonin, morphine+Met+N, and morphine+Met+ melatonin+N groups. On day 56 post-treatment, the blood was collected from the tail and the serum levels of sex hormones were evaluated, then the rats were sacrificed, and their bilateral testes and epididymis were retrieved for histological, immunohistochemical, molecular, testicular tissue stress oxidative status, and sperm parameters assays. Exposure to morphine, Met, and shift of morphine to Met resulted in testicular degeneration that can be attributed to generating the stress oxidative-induced- apoptotic testicular cell death and impairing spermatogenesis. Melatonin and N alone and particularly, in combination with each other improved testicular degeneration, sex hormone suppression, and testicular function mediated by increasing the testicular antioxidant capacity and inhibition of the apoptosis pathway. It's suggested that oral administration of antioxidants may be an effective treatment for attenuating some opioid-related testicular dysfunction and degeneration.

Demulsification of Crude Oil Emulsions Using Versatile and Eco-Friendly Demulsifiers Based on Cellulose Decorated with Imidazolium-Bearing Triazole Moiety.

Water in the crude oil forms emulsions that must be broken before refining processes. Here, the novel and versatile derivatives of imidazolium ionic liquids (ILs) containing triazole moiety [SIm-TazCn][Br] were synthesized via click chemistry and then grafted onto the cellulose (CEL) to prepare eco-friendly demulsifiers for breaking water-in-oil (W/O) emulsions. The prepared compounds with two kinds of alkyl chains named CEL[SIm-TazC6][Br] and CEL[SIm-TazC10][Br] were characterized and employed for demulsifying W/O (30:70 and 50:50 vol %) emulsions. Also, the effect of the type and concentration of each demulsifier on the phase separation and dehydration efficiency (DE) were investigated through the bottle test. According to the bottle test results, the CEL[SIm-TazC6][Br] derivative at 4000 ppm showed a good DE of 79% after 5 min, which increased to 82% after 24 h. The interfacial tension (IFT) of derivatives at different concentrations was measured. The minimum IFT value of 16.3 mN/m was obtained for CEL[SIm-TazC6][Br] with a shorter alkyl chain at 4000 ppm after 24 h. The green and efficient CEL-based surfactants significantly demulsified the W/O emulsions because of the collaboration between imidazolium and triazole moieties.

Combination of gamma irradiation and storage condition for improving mechanical and physical postharvest characteristics of fresh garlic cloves.

The aim of this study was the discrimination and optimization of irradiation effect under physical and mechanical experiments on garlic. The samples were irradiated with 0, 75, and 150 Gy doses and stored at 4 and 18°C for 5 months. Physical, mechanical, and color properties were measured in the period of storage. Based on the results, all irradiated garlic samples had less quality variation than control samples. Response surface methodology (RSM) optimized dose, storage time, and temperature of the stored garlic which was 75 Gy, 2 months, and 17°C, respectively. In addition, after finding the optimal dose, time, and temperature, the most effective factor as weight loss was obtained and the data were classified by the principal component analysis (PCA) approach. The results showed that the PCA method had a high ability to classify and separate the data obtained from measuring the physicochemical properties of garlic and cover 99% variance of data. Moreover, partial least square (PLS) was applied for predicting weight loss data with R2 0.9999. As well, a mechanical test was investigated for finding the best situation and duration of storage condition. Finally, irradiation prevented the destruction of garlic and saved garlic in the best quality as compared with control or nonirradiated samples. After all this, it can be decided to keep garlic in warehouses and transfer this product with minimum damage.

A fabricated hydrogel of hyaluronic acid/curcumin shows super-activity to heal the bacterial infected wound.

High risk of acute morbidities and even mortality from expanding the antibiotics resistant infectious wounds force indefinite efforts for development of high performance wound-healing materials. Herein, we design a procedure to fabricate a hyaluronic acid (HA)-based hydrogel to conjugate curcumin (Gel-H.P.Cur). The highlight of this work is to provide a favorite condition for capturing curcumin while protecting its structure and intensifying its activities because of the synchronization with HA. Accordingly, HA as a major component of dermis with a critical role in establishing skin health, could fortify the wound healing property as well as antibacterial activity of the hydrogel. Gel-H.P.Cur showed antibacterial properties against Pseudomonas aeruginosa (P. aeruginosa), which were examined by bactericidal efficiency, disk diffusion, anti-biofilm, and pyocyanin production assays. The effects of Gel-H.P.Cur on the inhibition of quorum sensing (QS) regulatory genes that contribute to expanding bacteria in the injured place was also significant. In addition, Gel-H.P.Cur showed high potential to heal the cutaneous wounds on the mouse excisional wound model with repairing histopathological damages rapidly and without scar. Taken together, the results strongly support Gel-H.P.Cur as a multipotent biomaterial for medical applications regarding the treatment of chronic, infected, and dehiscent wounds.

The protective effects of astaxanthin on pre-antral follicle degeneration in ovine vitrified/warmed ovarian tissue.

This study assesses the protective effects of astaxanthin (AST) against vitrification/warming-induced cryoinjuries of ovarian tissue slices in sheep. Cortical slices of slaughterhouse acquired-ovine ovaries were randomly distributed in different groups: fresh, toxicity, and five vitrification groups including vitrification in presence of 0 (control group), 1, 10 and 100 µM astaxanthin or 100 µM vitamin E. After vitrification/warming and 24 h culturing, the samples were subjected to histological studies, antioxidant evaluation by TAC and TBAR assays, and assessment of relative expression of BMP4, BMP15, GDF9 and KITLG genes related to folliculogenesis and follicular growth regulation. According to the results, vitrification reduced the percentage of morphologically intact follicles compared to the fresh and toxicity groups (p < 0.05). In vitrification groups, vitamin E and all three concentrations of AST increased the percentage of intact pre-antral follicles and antioxidant activity relative to the vitrified control (p < 0.05). This enhancement significantly occurred in 10 µM AST group more than vitamin E (p < 0.05). Also, 10 µM concentration of AST enhanced the expression of all the examined genes compared to the control (p < 0.05), while the expression of BMP4, BMP15 and KITLG was higher in the AST than vitamin E (p < 0.05). The latter could increase only the expression of GDF9 compared to the control group (p = 0.011). In conclusion, AST is a highly effective antioxidant for maintaining the survival of pre-antral follicles, retaining cell density, increasing total antioxidant capacity, and increasing the expression of some genes related to follicular development after short-term culture of vitrified/warmed ovarian tissue slices.

Application of imidazolium based ionic liquids grafted on microcrystalline cellulose as demulsifiers for water in crude oil (W/O) emulsions.

Separation of water and oil from water-in-oil (W/O) emulsion before transportation and refining is very important and critical. Ionic liquids (ILs) and their derivatives have recently attracted much attention as efficient chemical agents for breaking emulsions. So, here, a series of microcrystalline cellulose (MCC) grafted with imidazolium-based ionic liquids (IM-ILs) were synthesized, named as MCC@IM-ILs, and evaluated as eco-friendly surface-active agents for the separation of water from the crude oil. Structure of the synthesized compounds was confirmed by different characterization techniques. Dehydration efficiency percent (DE%) of the synthesized demulsifiers was measured and compared with each other. Synthesized MCC@IM-ILs showed an acceptable DE% to demulsify three kinds of W/O emulsions with different water content after 5 min. Concentration, alkyl chain length, and counter-anion of the synthesized MCC@IM-ILs play a key role in separating water from crude oil. Demulsifier with C10 alkyl chain length showed better DE% than the corresponding demulsifier with C6 alkyl chain length in the W/O (30:70 v/v) emulsion. Also, demulsifier with Br counter anion showed lower DE% than the corresponding BF4 ion-exchanged compound with higher hydrophilicity. Synthesized demulsifiers immobilized on ILs have significant advantages compared to unsupported ILs due to the use of green and economical cellulosic substrate.

Co-culturing or conditioned medium of Sertoli cells: Which one supports in vitro maturation of bovine oocytes and developmental competency of resulting embryos?

BACKGROUND: Sertoli cells (SCs) as supportive cells in the seminiferous tubule play an essential role in the nutrition and development of adjacent cells by secreting several beneficial growth factors, stimulators and cytokines which can be conceived to improve the developmental competency of oocyte or embryo in the co-culture system. OBJECTIVES: This study aimed to improve the maturation of bovine oocytes and consequently the development of resulting embryos in co-culture with SCs and their conditioned medium (CM). METHODS: The retrieved cumulus-oocyte complexes (COCs) from the abattoir-derived ovaries were matured in maturation medium alone (control group), in co-culture with ovine SCs (co-culture group), and in presence of 10% CM prepared in 33°C and 39°C (CM33 and CM39 groups). The nuclear maturation competency and subsequent embryo development rate of cultured COCs in all groups were evaluated. RESULTS: The results of this study showed that SCs and CM increased meiosis resumption from GV to the MII compared to the control group, significantly (p < 0.05). Besides, the degenerated oocytes in the co-culture group were significantly higher than those in the control, CM33 and CM39 groups (p < 0.05), and the lowest cleavage rate belonged to the co-culture group (p < 0.05). The blastocyst rate was also lower in the co-culture group than other groups and there was a significant difference between the control and two CM groups (p < 0.05). CONCLUSIONS: The Sertoli cells can be suitable for co-culturing with oocytes during IVM but detrimental for subsequent embryo development. In turn, Sertoli cell-derived conditioned medium (SC-CM) can provide sufficient bioactive materials for COCs to enhancing oocyte competence and embryo development.

The effect of quercetin in the maturation media on cumulus-granulosa cells and the developmental competence of bovine oocytes.

The present study was designed to investigate the effects of Quercetin on the developmental competence of bovine oocytes and cumulus-granulosa cells (CGs). Two groups of immature cumulus-oocyte complexes (COCs) were subjected to IVM with or without Quercetin. The viability, nuclear status, early and late apoptosis of oocytes and CGs were evaluated using gene expression analysis and staining methods. Embryonic development was assessed morphologically by recording Post-IVF survival, cleavage, and blastocyst rates. The proportion of oocytes reaching the MII stage was greater and the number of early-apoptotic oocytes was lower in the group matured with Quercetin compared to the Control (p < 0.05). Relative upregulation of OCT-4, IGF2R and Bcl-2, and downregulation of CHOP was seen in treated oocytes. Also, downregulation of Bax and upregulation of Glut-4 was detected in treated CGs. The treated oocytes experienced higher post-IVF survival and cleavage rates compared to the untreated group (p < 0.05); more cleaved embryos reached ≥16-cell stage and blastocyst at days 4th and 7th respectively. In addition, total blastocyst rate was significantly improved. It is concluded that supplementing maturation media with Quercetin can enhance the quality of bovine oocytes and endow them with protective potential against early apoptotic damage possibly through CHOP regulation of BCL2 gene family, triggering expression of a gene in CGs to maintain the intactness of oocyte against apoptotic signals and providing oocytes with more energy substrates. It also boosts the subsequent development of oocyte to blastocyst and improves the efficacy of bovine embryo production in vitro.

Biochemical and insecticidal effects of plant essential oils on insecticide resistant and susceptible populations of Musca domestica L. point to a potential cross-resistance risk.

Essential oils (EOs) can provide important alternatives to chemical insecticides in the control of pests. In this study, 12 EOs of native plant species from Iran were evaluated for their adulticidal activity against the house fly. In addition, we examined the insecticidal activity of Zataria multiflora and Rosmarinus officinalis EOs on adult female house flies from pyrethroid and organophosphate resistant and susceptible populations, using both fumigant and topical bioassays. The involvement of detoxification enzymes in susceptibility was investigated with synergism experiments in vivo, while the inhibitory effects of R. officinalis and Zataria multiflora EOs on the activities of cytochrome P450-dependent monooxygenases (P450s), carboxylesterases (CarEs) and glutathione S-transferases (GSTs) were determined by enzymatic inhibition assays in vitro. The EOs of Z. multiflora, Mentha pulegium, R. officinalis and Thymus vulgaris were the most effective against adults in contact topical assays, while oils extracted from Eucalyptus cinerea, Z. multiflora, Citrus sinensis, R. officinalis, Pinus eldarica and Lavandula angustifolia where the most effective in fumigant assays. Rosmarinus officinalis and Z. multiflora EOs were selected for further investigation and showed higher toxicity against a susceptible population, compared to two insecticide-resistant populations. Correlation analysis suggested cross-resistance between these EOs and pyrethroids in the resistant populations. The toxicity of both EOs on the resistant populations was synergized by three detoxification enzyme inhibitors. Further, in vitro inhibition studies showed that R. officinalis and Z. multiflora EOs more effectively inhibited the activities of the detoxification enzymes from flies of the susceptible population compared to those of the pyrethroid resistant populations. Synergistic and enzymatic assays further revealed that increased activities of P450s, GSTs, and CarEs are possibly involved in the cross-resistance between EOs and pyrethroids. Investigating the molecular mechanisms of P450s, GSTs, and CarEs in the resistance to EOs should be subject to further studies.

Ram epididymal sperm frozen in an extender containing ethylene glycol have higher post-thaw longevity and in vitro fertility.

BACKGROUND: Establishing an efficient, simple and inexpensive method for freezing ram epididymal sperm so that the quality and fertility of spermatozoa could be maintained for a longer period after thawing is of great practical value. OBJECTIVES: To optimize freezing and thawing protocol for ram epididymal sperm using either ethylene glycol (EG) or glycerol (GLY) as cryoprotectants (CPAs). Then, to evaluate the post-thaw longevity and in vitro fertility of spermatozoa that were frozen and thawed according to the optimized protocol. MATERIALS AND METHODS: At first, an optimum protocol for freezing and thawing sperm using EG or GLY were investigated, and the next experiments were performed using the spermatozoa that had been frozen and thawed according to the optimized protocol for each CPA. In the next experiments, frozen-thawed and fresh sperm were diluted in an isotonic culture medium and subsequently incubated at 39°C for 4 h. The motility characteristics and functional membrane integrity (FMI) of spermatozoa were evaluated after thawing, after dilution (t0 ), and after incubation (t4 ). The in vitro fertility of the spermatozoa was assessed at t0 and t4 . RESULTS: For both CPAs, the highest motility parameters and FMI was found for spermatozoa frozen at 3 cm above LN2 and thawed at 50 and 65°C (P < 0.05). In comparison to the spermatozoa of GLY group, the spermatozoa of the EG group had higher total and progressive motility at t0 , as well as higher FMI, total and progressive motility, and linearity at t4 (P < 0.05). Fertility of frozen-thawed sperm was higher than that of fresh sperm at t0 (P < 0.05). Incubation treatment increased the fertility of fresh sperm while decreased the fertility of frozen-thawed sperm, and this decline was more severe in GLY than in the EG group. CONCLUSION: Based on the findings, EG can be a more suitable CPA for freezing ram epididymal sperm.

The effect of chemical treatment of the sheep embryo zona pellucida on the ability of blastocysts to hatch after vitrification and warming.

BACKGROUND: The embryo release from the zona pellucida is of prerequisites of successful implantation. OBJECTIVES: Regarding the negative impact of embryo cryopreservation on the blastocysts hatchability, the aim of the present study was to investigate the effects of treating embryonic zona pellucida with pronase or acidic Tyrode's solution (ATS) before morula formation on the viability, freezability, and hatchability of vitrified-warmed resulted blastocysts. METHODS: In the first experiment, the zona pellucida of 3- and 4-day-old embryos were treated with the above compounds for 30 or 45 s. Then, the competency of the treated embryos to reach to blastocyst stage and the hatchability of resulting blastocysts were investigated. In the second experiment, the cryo-survivability and hatching rate of blastocysts resulting from 3-day-old embryos treated with pronase and ATS for 30 s were tested. RESULTS: In the first experiment and in contrast to the 45 s exposure, 30-s exposure of embryos to pronase or ATS did not have negative effect on the viability and development of embryos to blastocyst stage. In the second experiment, the freezability of blastocysts derived from 3-day-old embryos treated with pronase and ATS for 30 s was not different from that of the control group. However, the hatching rate of the pronase group was significantly higher than that of the control group. CONCLUSION: The results of the present study showed that reducing the thickness of zona pellucida of sheep embryos with pronase had no negative effect on the developmental competency and freezability of the treated embryos and improved the hatchability of vitrified-warmed blastocysts.

Human amniotic membrane stem cells' conditioned medium has better support for in-vitro production of bovine embryos than FBS.

Apart from oocyte quality, the media used has a significant effect on the production and quality of blastocysts produced in vitro. This study was designed to evaluate the replacement of serum with human amniotic membrane stem cells' conditioned medium (hAMSCs-CM) during bovine embryo culture on the quantity and quality of produced blastocysts. The in-vitro-produced embryos on the third day of IVC were randomly divided into the following culture groups: SOFaa + 5% FBS (Control), SOFaa + 5% hAMSCs-CM (5% CM), SOFaa + 2.5% hAMSCs-CM + 2.5% FBS (2.5% CM) and SOFaa + hAMSC co-culture (co-culture). The blastocyst and hatching rates, blastocyst cells number (the number of trophectoderm, inner cell mass and total cells), and the expression of some developmentally important genes (OCT4, PLAC8 and COX2 genes) in the treated groups, especially in the 5% CM, compared to the control had improved (p < .05). No significant difference was observed between groups for viability and hatching rate in vitrified-warmed blastocysts. Due to the positive effect of hAMSCs' conditioned medium (hAMSCs-CM) on blastocyst production, as well as its ease of preparation and the need to avoid the transmission of microbial contamination to the culture medium, hAMSCs-CM can be used as a suitable alternative to FBS during 3 to 8 days of bovine embryo culture.

Quercetin effect on the efficiency of ovine oocyte vitrification at GV stage.

The widely adopted method of vitrification is known to induce some negative effects on oocytes. In order to enhance the efficiency of this process performed on ovine oocytes at germinal vesicle stage, vitrification and warming (VW) solutions, and maturation media were supplemented with 5 µM Quercetin (Q). Four groups of vitrified and fresh immature oocytes were subjected to IVM, IVF and IVC, and their survival rate, apoptosis, nuclear status and developmental competence were assessed. Non-vitrified oocytes treated with Quercetin experienced higher cleavage rate relative to those matured without Quercetin (p < 0.05). Supplementation of VW and maturation media with Quercetin resulted in increased survival, cleavage and total blastocyst rates relative to the untreated oocytes. The post-IVM survival rate of non-vitrified oocytes showed no difference among those matured with and without Quercetin, but was higher for oocytes vitrified, warmed and matured with Quercetin relative to VW group lacking Quercetin. The proportion of early-apoptotic (AV+) oocytes was affected by Quercetin supplementation in both control and VW groups (p < 0.05). The number of AV positive oocytes was lower and the proportion of oocytes reaching MII stage was greater in non-vitrified and VW groups matured with Quercetin, in comparison with their untreated respective controls (p < 0.05). There was no difference in the number of late-stage apoptotic oocytes among different groups. It is concluded that supplementing vitrification and warming solutions with Quercetin endows vitrified ovine oocytes with protective potential against early apoptotic damage, and improves viability, maturation rate and developmental competence at GV stage.

A mesoporous melamine/chitosan/activated carbon biocomposite: Preparation, characterization and its application for Ni (II) uptake via ion imprinting.

A novel imprinted biocomposite and its non-imprinted form were developed by melaminating and crosslinking of chitosan coated onto a bio-based activated carbon and characterized using FTIR, BET, FESEM-EDS and XRD. Nickel, 4-Toluenesulfonyl chloride, and glutaraldehyde were used as a template, converter of hydroxyl and amine groups to good leaving groups, and cross-linker, respectively. The factors affecting adsorptivity and imprinting factor were optimized by using the Taguchi method for the subsequent comparative adsorptivity, kinetics, isotherms, selectivity, and reusability studies of imprinted biocomposite with its non-imprinted one. The pseudo-first-order and Langmuir models were best fitted to the experimental kinetics and equilibrium isotherm data, respectively. The maximum Ni (II)) adsorptivity of 109.86 mg/g, the imprinting factor (I·F) of 5.45 and Ni (II) selectivity coefficients values of 3.13, 4.48, 3.72, 2.51 for Ni (II) toward Zn (II), Cd (II), Cu (II) and Pb (II), respectively, were obtained at optimum conditions. After five consecutive adsorption-desorption cycles, the biocomposites still presented a high adsorptivity (>83%), indicating their excellent reusability.