Marwa Faheem - Academia.edu (original) (raw)

Papers by Marwa Faheem

Egyptian Journal of Animal Production, Apr 24, 2024

Theriogenology, 2016

Effects of α-tocopherol and freezing rates on the quality and heterologous in vitro fertilization... more Effects of α-tocopherol and freezing rates on the quality and heterologous in vitro fertilization capacity of stallion sperm after cryopreservation

Animal Reproduction Science, 2021

The physiological and molecular responses of granulosa cells (GCs) from buffalo follicles were in... more The physiological and molecular responses of granulosa cells (GCs) from buffalo follicles were investigated when there were in vitro heat stress conditions imposed. The cultured GCs were heat-treated at 40.5 °C for 24, 48 or 72 h while GCs of the control group were not heat-treated (37 °C). There were no differences in viability between control and heat-treated groups. There was an upward trend in increase in E2 secretion as the duration of heat stress advanced, being greater (P ≤ 0.05) for the GCs on which heat stress was imposed for 72 as compared with 24 h. In contrast, P4 release was less (P ≤ 0.05) from GCs heat-treated for 48 h than those cultured for 24 h and GCs of the control group. The relative abundance of ATP5F1A and SOD2 mRNA transcripts was consistent throughout the period when there was imposing of heat stress to sustain mitochondrial function. The relative abundance of CPT2 transcript was less in heat-treated GCs than in GCs of the control group. There was a greater relative abundance of SREBP1 and TNF-α mRNA transcripts after 48 h of heat-treatment of GCs than GCs of the control group. In conclusion, the results from the current study indicate buffalo GCs cultured when there was imposing of heat stress maintained normal viability, steroidogenesis and transcriptional profile. The stability of antioxidant status and increased transcription of genes regulating cholesterol biosynthesis and stress resistance may be defense mechanisms of buffalo GCs against heat stress.

Animals : an Open Access Journal from MDPI, 2021

Simple Summary The pertinent consequences of global warming substantially affect both animal prod... more Simple Summary The pertinent consequences of global warming substantially affect both animal productivity and fertility. Noteworthy, tropical and subtropical animal breeds are productively suited to hot climate conditions. Therefore studying the physiological changes accompanying high temperature, especially in tropically adapted species such as buffalo, will help in understanding the mechanisms that the animal use to accomplish the necessary functions efficiently. Concerning fertility-related activity, granulosa cells are important for the regulation of ovarian function and the completion of oocyte maturation. In this study, the buffalo granulosa cells were examined for their viability, physiological and molecular responses under in vitro heat stress conditions. Buffalo granulosa cells displayed different adaptive responses, at the physiological and molecular levels, to the different heat stress conditions. At 40.5 °C, granulosa cells exhibited a functional persistence compared to ...

Animals : an Open Access Journal from MDPI, 2021

Simple Summary The pertinent consequences of global warming substantially affect both animal prod... more Simple Summary The pertinent consequences of global warming substantially affect both animal productivity and fertility. Noteworthy, tropical and subtropical animal breeds are productively suited to hot climate conditions. Therefore studying the physiological changes accompanying high temperature, especially in tropically adapted species such as buffalo, will help in understanding the mechanisms that the animal use to accomplish the necessary functions efficiently. Concerning fertility-related activity, granulosa cells are important for the regulation of ovarian function and the completion of oocyte maturation. In this study, the buffalo granulosa cells were examined for their viability, physiological and molecular responses under in vitro heat stress conditions. Buffalo granulosa cells displayed different adaptive responses, at the physiological and molecular levels, to the different heat stress conditions. At 40.5 °C, granulosa cells exhibited a functional persistence compared to ...

Annals of Agricultural Science, Moshtohor, 2021

Effect ofheat shock and co-culture of monolayer granulosa cells (GCs) on in vitroembryo developme... more Effect ofheat shock and co-culture of monolayer granulosa cells (GCs) on in vitroembryo developmental competence in Egyptian buffalo was the aim of the present study.Cumulus oocytes complexes (COCs) were collected from ovaries (n= 448) of cyclicslaughtered buffalo cows. Good quality immature oocytes (n= 1512) were subjected to in vitro maturation and fertilization. Post 18-22 h of fertilization, presumptive embryos were randomly assigned into four groups: (G1) No heat shock (38.5 O C), (G2) Heat shock (40.5°C) and (G3) Co-culture with GCs monolayer and heat shock or (G4) Co-culture with GCs monolayer and no heat shock. Heatshockedembryos were exposed to temperature of40.5°Cfor the first two hours of culture before exposing to 38.5°C afterward up to the end of culture perio d (8 days). The data were analyzed by General LinearUnivariate model using SPSS. Oocyte expansionrate was 90.8±1.1%. This percentage is higher than nuclear maturation rate (oocytes at Telophase and Metaphase II) as detected byaceto-orcein staining by about 17%. Cleavage rate as recorded at day 3 post fertilization (day 0) was higher (p≤0.05) for G1 and G4 (71.1±10.5% and 70.5±7.9 with GCs, respectively).On the other hand, cleavage rate was dramatically decreased (P ≤0.05) for embryos of G2 (43.7±7.0%) compared with that of G3 (80.2±7.0%). In addition, embryos of G3showed approximately the same rate ofdeveloped embryos(Morula and blastocyst stages at day 8 of culture) as of G1 (50.9±5.3 and51.7±7.9%, respectively). In conclusion, using co-culture of GCs as a monolayer enhances cleavage rate and subsequent embryo development of buffalo embryos under heat stress condition.

Journal of World's Poultry Research, 2020

The current study was conducted to investigate the possible protective effect of curcumin supplem... more The current study was conducted to investigate the possible protective effect of curcumin supplementation on buffalo granulosa cells (GCs) under in vitro culture condition. Buffalo ovaries were collected from local abattoir in physiological saline solution and transported directly to laboratory. Follicular fluid containing GCs and cumulusoocyte-complexes were aspirated from antral follicles with diameter 2-8 mm. The collected GCs were seeded (Approximately 375,000 viable cells) in an 8-well culture plate containing tissue culture medium-199 (TCM-199) and kept at 37 °C in a humidified atmosphere of 5% CO 2. The curcumin was supplemented to TCM media at levels of 1, 2.5, 5 and 10 μM for 24 and 48 h at 37 °C or kept without treatment as control group. The viability of cells was determined using the trypan blue test. Intracellular reactive oxygen species (ROS) level was assessed by measuring the fluorescent intensity of 6-carboxy-2′,7′-dichlorodihydro fluorescein diacetate (H 2 DCFDA). In addition, mitochondrial activity of GCs was determined. The results of the present study indicated that the viability of GCs under culture conditions was significantly decreased in groups treated with 1, 2.5, 5 and 10 µM curcumin (86.0%, 86.26%, 83.0% and 74.0%, respectively) compared to control group (93.60 %). The two groups of granulosa cells cultured with 2.5 and 5 µM curcumin recorded greater level of mitochondrial activity than the groups cultured with 1 µM and 10 µM curcumin. Moreover, there was a significant increase in ROS level in group cultured with 10 µM curcumin, compared to control and other experimental groups. The enzyme activity of catalase (CAT), superoxide dismutase (SOD), glutathione (GSH) and 1, 1-diphenyl-2-picrylhydrazyl (DPPH) was increased after treating in vitro cultured granulosa cells with 5 µM of curcumin. However, the enzymatic activity of CAT, SOD, GSH and DPPH was declined significantly 48 h post-curcumin treatment. In conclusion, supplementation of curcumin at low concentration (2.5 µM) for 24 h to in vitro cultured GCs improved intracellular metabolic activity and antioxidant protective system, whereas it could not sustain this action for 48 h. Moreover, supplementation of curcumin at high concentration and for long duration may negatively affect viability of GCs under in vitro culture condition via induction of oxidative stress.

Journal of World's Poultry Research, 2020

The aim of this investigation was to follow up in vitro preimplantation development of buffalo cu... more The aim of this investigation was to follow up in vitro preimplantation development of buffalo cumulus-oocyte complexes (COCs) after BCB test and followed by in vitro maturation under two different levels of oxygen tension. Cumulus-oocyte complexes (n=1045) were selected with BCB staining (oocytes with any degree of blue color in cytoplasm was defined as BCB+, oocytes without any degree of blue color in cytoplasm was defined as BCB-) in addition to a third control group. The previous experimental groups (BCB+, BCB-, control) were matured in vitro under low (5%) and high oxygen tension (20%), followed by in vitro fertilization and in vitro culture of presumptive zygotes. There were no differences (P ≤ 0.05) in cleavage, morula and transferable embryos rates among BCB+, BCB-and control group. However, blastocyst rate was greater significantly in control group (14.4 ± 2.0) than BCB-COCs (8.4 ± 1.9). According to the oxygen tension effect, the rate of morula and transferable embryos was increased (P ≤ 0.05) in buffalo COCs developed under low oxygen tension (11.6 ± 1.4 and 23.8 ± 1.9) compared to high oxygen tension group (7.4 ± 1.4 and 17.9 ± 2.1). In addition, cleavage, morula, blastocyst and transferable embryos rates were greater in BCB+ under low (43.6 ± 3.9, 14.9 ± 2.5, 14.1 ± 2.9 and 28.4 ± 3.6) than high oxygen tension group (33.5 ± 3.9, 7.1 ± 2.5, 11.6 ± 2.9 and 18.8 ± 3.6) which may reflect enhanced biological processes controlling early development. Moreover, blastocyst rate was significantly higher in control group cultured under high (12.0 ± 2.9) and low (16.9 ± 2.8) oxygen level than their counterparts of BCB-group (9.3 ± 2.9 and 7.6 ± 2.6, respectively). In conclusion, there was no differences in embryo development between BCB+ and BCB-, COCs; therefore, oocyte selection based on BCB staining is not an effective tool to select developmental competent buffalo COCs. Buffalo morula and transferable embryos prefer low oxygen tension for early development, which should be applied during in vitro embryo production of this species.

Animal Reproduction Science, 2021

The physiological and molecular responses of granulosa cells (GCs) from buffalo follicles were in... more The physiological and molecular responses of granulosa cells (GCs) from buffalo follicles were investigated when there were in vitro heat stress conditions imposed. The cultured GCs were heat-treated at 40.5 °C for 24, 48 or 72 h while GCs of the control group were not heat-treated (37 °C). There were no differences in viability between control and heat-treated groups. There was an upward trend in increase in E2 secretion as the duration of heat stress advanced, being greater (P ≤ 0.05) for the GCs on which heat stress was imposed for 72 as compared with 24 h. In contrast, P4 release was less (P ≤ 0.05) from GCs heat-treated for 48 h than those cultured for 24 h and GCs of the control group. The relative abundance of ATP5F1A and SOD2 mRNA transcripts was consistent throughout the period when there was imposing of heat stress to sustain mitochondrial function. The relative abundance of CPT2 transcript was less in heat-treated GCs than in GCs of the control group. There was a greater relative abundance of SREBP1 and TNF-α mRNA transcripts after 48 h of heat-treatment of GCs than GCs of the control group. In conclusion, the results from the current study indicate buffalo GCs cultured when there was imposing of heat stress maintained normal viability, steroidogenesis and transcriptional profile. The stability of antioxidant status and increased transcription of genes regulating cholesterol biosynthesis and stress resistance may be defense mechanisms of buffalo GCs against heat stress.

Animals : an Open Access Journal from MDPI, 2021

Simple Summary The pertinent consequences of global warming substantially affect both animal prod... more Simple Summary The pertinent consequences of global warming substantially affect both animal productivity and fertility. Noteworthy, tropical and subtropical animal breeds are productively suited to hot climate conditions. Therefore studying the physiological changes accompanying high temperature, especially in tropically adapted species such as buffalo, will help in understanding the mechanisms that the animal use to accomplish the necessary functions efficiently. Concerning fertility-related activity, granulosa cells are important for the regulation of ovarian function and the completion of oocyte maturation. In this study, the buffalo granulosa cells were examined for their viability, physiological and molecular responses under in vitro heat stress conditions. Buffalo granulosa cells displayed different adaptive responses, at the physiological and molecular levels, to the different heat stress conditions. At 40.5 °C, granulosa cells exhibited a functional persistence compared to ...

Egyptian Journal of Animal Production, 2018

The current study was designed to investigate the expression profile of selected candidate genes ... more The current study was designed to investigate the expression profile of selected candidate genes in corpus luteum of cyclic Egyptian buffaloes. Ovaries were collected from 18 cyclic females slaughtered in Egyptian local a battoir. Corpus luteum stages of development (growth: around days 3-5, static: around days 7-10 and regression: around days 19-21 of estrous cycle) were determined by morphological features and based on our previous studiesd one in this species. Quantitative realtime polymerase chain reaction (PCR) was performed to profile the transcript abundance of genes involved in mitochondrial activity (SOD2, TFAM and CPT2), lipid metabolism (SREBP1), proinflammatory cytokine (TNFα), oxidative stress (NFE2L2),while GAPDH was used as housekeeping gene. The relative gene expression profiles of TFAM and SOD2 were up-regulated significantly (P≤0.05) in static compared to both growing and regressed phases. Meanwhile, the transcript abundance of both CPT2 and SREBP1 was significantl...

Zygote, 2021

SummaryThis study was conducted to monitor the cellular and molecular changes of buffalo cumulus–... more SummaryThis study was conducted to monitor the cellular and molecular changes of buffalo cumulus–oocytes complexes (COCs) cultured under high or low oxygen levels. Morphologically good quality COCs (n = 1627) were screened using brilliant cresyl blue (BCB) staining and placed into three groups (BCB+, BCB− and control). All groups of COCs were cultured under low (5%) or high (20%) oxygen tensions. Intracellular and molecular changes including oocyte ultrastructure, lipid contents, mitochondrial activity and transcript abundance of genes regulating different pathways were analyzed in the matured oocyte groups. The results revealed that oxygen tension did not affect cumulus expansion rates, however the BCB+ group had a higher (P ≤ 0.05) expansion rate compared with the BCB− group. BCB− oocytes recorded the lowest meiotic progression rate (P ≤ 0.05) under high oxygen levels that was linked with an increased level of reactive oxygen species (ROS) compared with the BCB+ oocytes. Ultrastru...

Egyptian Journal of Animal Production, 2018

The current study was designed to investigate the expression profile of selected candidate genes ... more The current study was designed to investigate the expression profile of selected candidate genes in corpus luteum of cyclic Egyptian buffaloes. Ovaries were collected from 18 cyclic females slaughtered in Egyptian local a battoir. Corpus luteum stages of development (growth: around days 3-5, static: around days 7-10 and regression: around days 19-21 of estrous cycle) were determined by morphological features and based on our previous studiesd one in this species. Quantitative realtime polymerase chain reaction (PCR) was performed to profile the transcript abundance of genes involved in mitochondrial activity (SOD2, TFAM and CPT2), lipid metabolism (SREBP1), proinflammatory cytokine (TNFα), oxidative stress (NFE2L2),while GAPDH was used as housekeeping gene. The relative gene expression profiles of TFAM and SOD2 were up-regulated significantly (P≤0.05) in static compared to both growing and regressed phases. Meanwhile, the transcript abundance of both CPT2 and SREBP1 was significantl...

Egypt has the largest buffalo population in the Middle East and Africa (3.717 million); they act ... more Egypt has the largest buffalo population in the Middle East and Africa (3.717 million); they act as a major source of red meat as well as the major producer of milk with a high efficiency of converting poor quality roughage into meat and milk. In addition, buffaloes are inherently resistant to many climatic stresses. However buffaloes are reported to have low reproductive performance with inherent reproductive problems of weak/silent estrus signs, seasonal anestrous. So, attention has been towards to biotechnologies as a way for overcome these problems and moreover to augment the reproduction potential of these animals. The present book is provide a detailed research on in vitro production of buffalo embryos, studying extensively the changes occurred in the oocyte morphology and morphometry during oocyte maturation, the stages of embryonic development after fertilization and the developmental kinetics of the in vitro produced embryos. This book” In vitro buffalo embryonic developmen...

Egyptian Journal of Animal Production, 2007

Two experiments (Exp) were carried out to test the effect of different media on in vitro maturati... more Two experiments (Exp) were carried out to test the effect of different media on in vitro maturation (IVM) (Exp 1; n = 1142 ovaries with 2666 oocytes) and to study in vitro fertilization (IVF) of buffalo oocytes (Exp 2; n = 110 ovaries with 330 oocytes). In exp 1 the collected cumulus oocyte complex (COCs) was allocated to three maturational media; TCM-199 supplemented with hormones (M1), TCM-199 supplemented with epidermal growth factor (M2) and m-SOF (M3) overlaid with paraffin oil before incubating under 5 % CO 2 in air at 38.5 o C. A part of COCs was fixed to determine the stages of nuclear maturation after 24 hr. In exp 2 the matured oocytes were allotted with sperm suspension before incubating under 5 % CO 2 in air at 38.5 o C, for 22-24 hr. Presumptive embryos were placed, individually in 96 wells petri dish to determine at 12 hr intervals the kinetics of cleavage up to blastocyst stage. Average of collected COCs per ovary was 2.4 with 86 % of excellent and good graded COC's. M1 and M2 showed higher (P<0.05) maturation rate vs. M3. After culturing oocyte diameter and perivitalline space increased (P<0.05) relative to preculture diameters, while diameter of ooplasm and thickness of zona pellucida non significantly decreased. Fertilized ova and cleaved embryos (%) were 80.2 and 77.8 %, respectively. Early embryonic cleavage up to 32 cell stage lapsed took 84 hr, while corresponding periods to reach morula, and blastocyst stage were 101 and 134.5 hr, respectively. Out of the fertilized ova 31.5 % reached excellent and good graded blastocyst stage.

Egyptian Journal of Animal Production, 2019

The current study was done to investigate the expression profile of selected candidate genes in c... more The current study was done to investigate the expression profile of selected candidate genes in corpus luteum of cyclic Egyptian buffaloes. A total number of nine corpora lutea (CLs) was collected from 9 cyclic slaughtered female Egyptian buffaloes. Based on their developmental phases and morphological features, CLs were classified into; growing (around days 3-5), static (around days 7-10) and regressed (around days 19-21 of the estrous cycle). Transcript abundance of genes regulating mitochondrial activity (SOD2, TFAM and CPT2), lipid metabolism (SREBP1), pro-inflammatory cytokine (TNFα) and oxidative stress (NFE2L2) was assessed relative to GAPDH as a housekeeping gene. Relative gene expression profiles of TFAM and SOD2 increased (P ≤ 0.05) in static phase compared to both growing and regressed phases. Transcript abundance of both CPT2 and SREBP1 was significantly higher at static phase, while was at minimum level of expression during regression phase. The expression profile of NFE2L2 revealed insignificant differences among the studied phases of CLs. Relative transcript abundance of TNFα was higher (P ≤ 0.05) at regression phase compared to growing and static ones. Taken together, the pattern of genes regulating mitochondrial activity and lipid metabolism were higher during growing phase and reached a plateau in static phase and fully declined during regression phase. Meanwhile, the transcript abundance of pro-inflammatory cytokine gene denoted an opposite trend. Thus, gene expression profile reflected the cyclic changes of buffalo corpus luteum development.

Canadian Journal of Animal Science, 2016

Feeding ruminants with Pittosporum undulatum can adversely affect the immune, endocrine and repro... more Feeding ruminants with Pittosporum undulatum can adversely affect the immune, endocrine and reproductive systems. The aim of the present study was to evaluate how feeding with P. undulatum alters progesterone levels during the oestrus cycle, and fertilization and development of oocytes after in vitro fertilization (IVF). Heifers were fed a diet containing (n=4) or not containing (n=4) this plant. Peripheral blood samples were collected every three days and progesterone levels were analysed by ELISA. The heifers' ovaries were recovered after slaughter and the oocytes were removed, maturated, in vitro fertilized and cultured for seven days, during which time their development was evaluated every two days. Results indicated that plasma concentrations of progesterone during the oestrus cycle were significantly higher (p <0.01) in the luteal and follicular phases of the animals fed without Pittosporum when compared with those fed with Pittosporum. Statistical differences (p <0.05) in maturation, cleavage rate and embryonic development were observed between oocytes belonging to the experimental and control group. Rates of maturation, fertilisation and embryo development in the experimental group were, respectively, 78.3±5.8, 29.92±4.31 and 7.30±3.1 as compared to 90.5±3.0, 41.86±58 and 21.88±6.85 in the control group. These differences can be explained as compounds from this plant can directly or indirectly inactivate cyclooxygenase 2 (COX-2), which may diminish follicular development through the inhibition of prostaglandins synthesis, oocyte maturation and, consequently, reduce the ability of oocytes to be fertilized and develop.

Zygote (Cambridge, England), Jan 8, 2017

The exposure of oocytes to heat stress during the maturation process results in harmful effects t... more The exposure of oocytes to heat stress during the maturation process results in harmful effects to their internal organelles, low fertilization capability and higher embryonic losses. In the present experiment the effect of heat shock (HS) during the maturation process was assessed. In Assay 1, oocytes from winter (December-March; n = 100) and summer (June-September; n = 100) months were collected and matured to analyse their HS tolerance. Total RNA was extracted from matured oocytes and cDNA synthesis was performed, followed by qPCR for selected genes (Cx43, CDH1, DNMT1, HSPA14), compared with two reference genes (GAPDH and SDHA). In Assay 2, oocytes collected during the winter were subjected to kinetic HS by stressing them at 39.5°C for 6, 12, 18 or 24 h and were afterwards matured at control temperature (38.5°C), and subsequently subjected to the previously described gene analysis procedure. Results of Assay 1 show that summer-collected oocytes exhibited lower maturation rate tha...

Egyptian Journal of Animal Production, Apr 24, 2024

Theriogenology, 2016

Effects of α-tocopherol and freezing rates on the quality and heterologous in vitro fertilization... more Effects of α-tocopherol and freezing rates on the quality and heterologous in vitro fertilization capacity of stallion sperm after cryopreservation

Animal Reproduction Science, 2021

The physiological and molecular responses of granulosa cells (GCs) from buffalo follicles were in... more The physiological and molecular responses of granulosa cells (GCs) from buffalo follicles were investigated when there were in vitro heat stress conditions imposed. The cultured GCs were heat-treated at 40.5 °C for 24, 48 or 72 h while GCs of the control group were not heat-treated (37 °C). There were no differences in viability between control and heat-treated groups. There was an upward trend in increase in E2 secretion as the duration of heat stress advanced, being greater (P ≤ 0.05) for the GCs on which heat stress was imposed for 72 as compared with 24 h. In contrast, P4 release was less (P ≤ 0.05) from GCs heat-treated for 48 h than those cultured for 24 h and GCs of the control group. The relative abundance of ATP5F1A and SOD2 mRNA transcripts was consistent throughout the period when there was imposing of heat stress to sustain mitochondrial function. The relative abundance of CPT2 transcript was less in heat-treated GCs than in GCs of the control group. There was a greater relative abundance of SREBP1 and TNF-α mRNA transcripts after 48 h of heat-treatment of GCs than GCs of the control group. In conclusion, the results from the current study indicate buffalo GCs cultured when there was imposing of heat stress maintained normal viability, steroidogenesis and transcriptional profile. The stability of antioxidant status and increased transcription of genes regulating cholesterol biosynthesis and stress resistance may be defense mechanisms of buffalo GCs against heat stress.

Animals : an Open Access Journal from MDPI, 2021

Simple Summary The pertinent consequences of global warming substantially affect both animal prod... more Simple Summary The pertinent consequences of global warming substantially affect both animal productivity and fertility. Noteworthy, tropical and subtropical animal breeds are productively suited to hot climate conditions. Therefore studying the physiological changes accompanying high temperature, especially in tropically adapted species such as buffalo, will help in understanding the mechanisms that the animal use to accomplish the necessary functions efficiently. Concerning fertility-related activity, granulosa cells are important for the regulation of ovarian function and the completion of oocyte maturation. In this study, the buffalo granulosa cells were examined for their viability, physiological and molecular responses under in vitro heat stress conditions. Buffalo granulosa cells displayed different adaptive responses, at the physiological and molecular levels, to the different heat stress conditions. At 40.5 °C, granulosa cells exhibited a functional persistence compared to ...

Animals : an Open Access Journal from MDPI, 2021

Simple Summary The pertinent consequences of global warming substantially affect both animal prod... more Simple Summary The pertinent consequences of global warming substantially affect both animal productivity and fertility. Noteworthy, tropical and subtropical animal breeds are productively suited to hot climate conditions. Therefore studying the physiological changes accompanying high temperature, especially in tropically adapted species such as buffalo, will help in understanding the mechanisms that the animal use to accomplish the necessary functions efficiently. Concerning fertility-related activity, granulosa cells are important for the regulation of ovarian function and the completion of oocyte maturation. In this study, the buffalo granulosa cells were examined for their viability, physiological and molecular responses under in vitro heat stress conditions. Buffalo granulosa cells displayed different adaptive responses, at the physiological and molecular levels, to the different heat stress conditions. At 40.5 °C, granulosa cells exhibited a functional persistence compared to ...

Annals of Agricultural Science, Moshtohor, 2021

Effect ofheat shock and co-culture of monolayer granulosa cells (GCs) on in vitroembryo developme... more Effect ofheat shock and co-culture of monolayer granulosa cells (GCs) on in vitroembryo developmental competence in Egyptian buffalo was the aim of the present study.Cumulus oocytes complexes (COCs) were collected from ovaries (n= 448) of cyclicslaughtered buffalo cows. Good quality immature oocytes (n= 1512) were subjected to in vitro maturation and fertilization. Post 18-22 h of fertilization, presumptive embryos were randomly assigned into four groups: (G1) No heat shock (38.5 O C), (G2) Heat shock (40.5°C) and (G3) Co-culture with GCs monolayer and heat shock or (G4) Co-culture with GCs monolayer and no heat shock. Heatshockedembryos were exposed to temperature of40.5°Cfor the first two hours of culture before exposing to 38.5°C afterward up to the end of culture perio d (8 days). The data were analyzed by General LinearUnivariate model using SPSS. Oocyte expansionrate was 90.8±1.1%. This percentage is higher than nuclear maturation rate (oocytes at Telophase and Metaphase II) as detected byaceto-orcein staining by about 17%. Cleavage rate as recorded at day 3 post fertilization (day 0) was higher (p≤0.05) for G1 and G4 (71.1±10.5% and 70.5±7.9 with GCs, respectively).On the other hand, cleavage rate was dramatically decreased (P ≤0.05) for embryos of G2 (43.7±7.0%) compared with that of G3 (80.2±7.0%). In addition, embryos of G3showed approximately the same rate ofdeveloped embryos(Morula and blastocyst stages at day 8 of culture) as of G1 (50.9±5.3 and51.7±7.9%, respectively). In conclusion, using co-culture of GCs as a monolayer enhances cleavage rate and subsequent embryo development of buffalo embryos under heat stress condition.

Journal of World's Poultry Research, 2020

The current study was conducted to investigate the possible protective effect of curcumin supplem... more The current study was conducted to investigate the possible protective effect of curcumin supplementation on buffalo granulosa cells (GCs) under in vitro culture condition. Buffalo ovaries were collected from local abattoir in physiological saline solution and transported directly to laboratory. Follicular fluid containing GCs and cumulusoocyte-complexes were aspirated from antral follicles with diameter 2-8 mm. The collected GCs were seeded (Approximately 375,000 viable cells) in an 8-well culture plate containing tissue culture medium-199 (TCM-199) and kept at 37 °C in a humidified atmosphere of 5% CO 2. The curcumin was supplemented to TCM media at levels of 1, 2.5, 5 and 10 μM for 24 and 48 h at 37 °C or kept without treatment as control group. The viability of cells was determined using the trypan blue test. Intracellular reactive oxygen species (ROS) level was assessed by measuring the fluorescent intensity of 6-carboxy-2′,7′-dichlorodihydro fluorescein diacetate (H 2 DCFDA). In addition, mitochondrial activity of GCs was determined. The results of the present study indicated that the viability of GCs under culture conditions was significantly decreased in groups treated with 1, 2.5, 5 and 10 µM curcumin (86.0%, 86.26%, 83.0% and 74.0%, respectively) compared to control group (93.60 %). The two groups of granulosa cells cultured with 2.5 and 5 µM curcumin recorded greater level of mitochondrial activity than the groups cultured with 1 µM and 10 µM curcumin. Moreover, there was a significant increase in ROS level in group cultured with 10 µM curcumin, compared to control and other experimental groups. The enzyme activity of catalase (CAT), superoxide dismutase (SOD), glutathione (GSH) and 1, 1-diphenyl-2-picrylhydrazyl (DPPH) was increased after treating in vitro cultured granulosa cells with 5 µM of curcumin. However, the enzymatic activity of CAT, SOD, GSH and DPPH was declined significantly 48 h post-curcumin treatment. In conclusion, supplementation of curcumin at low concentration (2.5 µM) for 24 h to in vitro cultured GCs improved intracellular metabolic activity and antioxidant protective system, whereas it could not sustain this action for 48 h. Moreover, supplementation of curcumin at high concentration and for long duration may negatively affect viability of GCs under in vitro culture condition via induction of oxidative stress.

Journal of World's Poultry Research, 2020

The aim of this investigation was to follow up in vitro preimplantation development of buffalo cu... more The aim of this investigation was to follow up in vitro preimplantation development of buffalo cumulus-oocyte complexes (COCs) after BCB test and followed by in vitro maturation under two different levels of oxygen tension. Cumulus-oocyte complexes (n=1045) were selected with BCB staining (oocytes with any degree of blue color in cytoplasm was defined as BCB+, oocytes without any degree of blue color in cytoplasm was defined as BCB-) in addition to a third control group. The previous experimental groups (BCB+, BCB-, control) were matured in vitro under low (5%) and high oxygen tension (20%), followed by in vitro fertilization and in vitro culture of presumptive zygotes. There were no differences (P ≤ 0.05) in cleavage, morula and transferable embryos rates among BCB+, BCB-and control group. However, blastocyst rate was greater significantly in control group (14.4 ± 2.0) than BCB-COCs (8.4 ± 1.9). According to the oxygen tension effect, the rate of morula and transferable embryos was increased (P ≤ 0.05) in buffalo COCs developed under low oxygen tension (11.6 ± 1.4 and 23.8 ± 1.9) compared to high oxygen tension group (7.4 ± 1.4 and 17.9 ± 2.1). In addition, cleavage, morula, blastocyst and transferable embryos rates were greater in BCB+ under low (43.6 ± 3.9, 14.9 ± 2.5, 14.1 ± 2.9 and 28.4 ± 3.6) than high oxygen tension group (33.5 ± 3.9, 7.1 ± 2.5, 11.6 ± 2.9 and 18.8 ± 3.6) which may reflect enhanced biological processes controlling early development. Moreover, blastocyst rate was significantly higher in control group cultured under high (12.0 ± 2.9) and low (16.9 ± 2.8) oxygen level than their counterparts of BCB-group (9.3 ± 2.9 and 7.6 ± 2.6, respectively). In conclusion, there was no differences in embryo development between BCB+ and BCB-, COCs; therefore, oocyte selection based on BCB staining is not an effective tool to select developmental competent buffalo COCs. Buffalo morula and transferable embryos prefer low oxygen tension for early development, which should be applied during in vitro embryo production of this species.

Animal Reproduction Science, 2021

The physiological and molecular responses of granulosa cells (GCs) from buffalo follicles were in... more The physiological and molecular responses of granulosa cells (GCs) from buffalo follicles were investigated when there were in vitro heat stress conditions imposed. The cultured GCs were heat-treated at 40.5 °C for 24, 48 or 72 h while GCs of the control group were not heat-treated (37 °C). There were no differences in viability between control and heat-treated groups. There was an upward trend in increase in E2 secretion as the duration of heat stress advanced, being greater (P ≤ 0.05) for the GCs on which heat stress was imposed for 72 as compared with 24 h. In contrast, P4 release was less (P ≤ 0.05) from GCs heat-treated for 48 h than those cultured for 24 h and GCs of the control group. The relative abundance of ATP5F1A and SOD2 mRNA transcripts was consistent throughout the period when there was imposing of heat stress to sustain mitochondrial function. The relative abundance of CPT2 transcript was less in heat-treated GCs than in GCs of the control group. There was a greater relative abundance of SREBP1 and TNF-α mRNA transcripts after 48 h of heat-treatment of GCs than GCs of the control group. In conclusion, the results from the current study indicate buffalo GCs cultured when there was imposing of heat stress maintained normal viability, steroidogenesis and transcriptional profile. The stability of antioxidant status and increased transcription of genes regulating cholesterol biosynthesis and stress resistance may be defense mechanisms of buffalo GCs against heat stress.

Animals : an Open Access Journal from MDPI, 2021

Simple Summary The pertinent consequences of global warming substantially affect both animal prod... more Simple Summary The pertinent consequences of global warming substantially affect both animal productivity and fertility. Noteworthy, tropical and subtropical animal breeds are productively suited to hot climate conditions. Therefore studying the physiological changes accompanying high temperature, especially in tropically adapted species such as buffalo, will help in understanding the mechanisms that the animal use to accomplish the necessary functions efficiently. Concerning fertility-related activity, granulosa cells are important for the regulation of ovarian function and the completion of oocyte maturation. In this study, the buffalo granulosa cells were examined for their viability, physiological and molecular responses under in vitro heat stress conditions. Buffalo granulosa cells displayed different adaptive responses, at the physiological and molecular levels, to the different heat stress conditions. At 40.5 °C, granulosa cells exhibited a functional persistence compared to ...

Egyptian Journal of Animal Production, 2018

The current study was designed to investigate the expression profile of selected candidate genes ... more The current study was designed to investigate the expression profile of selected candidate genes in corpus luteum of cyclic Egyptian buffaloes. Ovaries were collected from 18 cyclic females slaughtered in Egyptian local a battoir. Corpus luteum stages of development (growth: around days 3-5, static: around days 7-10 and regression: around days 19-21 of estrous cycle) were determined by morphological features and based on our previous studiesd one in this species. Quantitative realtime polymerase chain reaction (PCR) was performed to profile the transcript abundance of genes involved in mitochondrial activity (SOD2, TFAM and CPT2), lipid metabolism (SREBP1), proinflammatory cytokine (TNFα), oxidative stress (NFE2L2),while GAPDH was used as housekeeping gene. The relative gene expression profiles of TFAM and SOD2 were up-regulated significantly (P≤0.05) in static compared to both growing and regressed phases. Meanwhile, the transcript abundance of both CPT2 and SREBP1 was significantl...

Zygote, 2021

SummaryThis study was conducted to monitor the cellular and molecular changes of buffalo cumulus–... more SummaryThis study was conducted to monitor the cellular and molecular changes of buffalo cumulus–oocytes complexes (COCs) cultured under high or low oxygen levels. Morphologically good quality COCs (n = 1627) were screened using brilliant cresyl blue (BCB) staining and placed into three groups (BCB+, BCB− and control). All groups of COCs were cultured under low (5%) or high (20%) oxygen tensions. Intracellular and molecular changes including oocyte ultrastructure, lipid contents, mitochondrial activity and transcript abundance of genes regulating different pathways were analyzed in the matured oocyte groups. The results revealed that oxygen tension did not affect cumulus expansion rates, however the BCB+ group had a higher (P ≤ 0.05) expansion rate compared with the BCB− group. BCB− oocytes recorded the lowest meiotic progression rate (P ≤ 0.05) under high oxygen levels that was linked with an increased level of reactive oxygen species (ROS) compared with the BCB+ oocytes. Ultrastru...

Egyptian Journal of Animal Production, 2018

The current study was designed to investigate the expression profile of selected candidate genes ... more The current study was designed to investigate the expression profile of selected candidate genes in corpus luteum of cyclic Egyptian buffaloes. Ovaries were collected from 18 cyclic females slaughtered in Egyptian local a battoir. Corpus luteum stages of development (growth: around days 3-5, static: around days 7-10 and regression: around days 19-21 of estrous cycle) were determined by morphological features and based on our previous studiesd one in this species. Quantitative realtime polymerase chain reaction (PCR) was performed to profile the transcript abundance of genes involved in mitochondrial activity (SOD2, TFAM and CPT2), lipid metabolism (SREBP1), proinflammatory cytokine (TNFα), oxidative stress (NFE2L2),while GAPDH was used as housekeeping gene. The relative gene expression profiles of TFAM and SOD2 were up-regulated significantly (P≤0.05) in static compared to both growing and regressed phases. Meanwhile, the transcript abundance of both CPT2 and SREBP1 was significantl...

Egypt has the largest buffalo population in the Middle East and Africa (3.717 million); they act ... more Egypt has the largest buffalo population in the Middle East and Africa (3.717 million); they act as a major source of red meat as well as the major producer of milk with a high efficiency of converting poor quality roughage into meat and milk. In addition, buffaloes are inherently resistant to many climatic stresses. However buffaloes are reported to have low reproductive performance with inherent reproductive problems of weak/silent estrus signs, seasonal anestrous. So, attention has been towards to biotechnologies as a way for overcome these problems and moreover to augment the reproduction potential of these animals. The present book is provide a detailed research on in vitro production of buffalo embryos, studying extensively the changes occurred in the oocyte morphology and morphometry during oocyte maturation, the stages of embryonic development after fertilization and the developmental kinetics of the in vitro produced embryos. This book” In vitro buffalo embryonic developmen...

Egyptian Journal of Animal Production, 2007

Two experiments (Exp) were carried out to test the effect of different media on in vitro maturati... more Two experiments (Exp) were carried out to test the effect of different media on in vitro maturation (IVM) (Exp 1; n = 1142 ovaries with 2666 oocytes) and to study in vitro fertilization (IVF) of buffalo oocytes (Exp 2; n = 110 ovaries with 330 oocytes). In exp 1 the collected cumulus oocyte complex (COCs) was allocated to three maturational media; TCM-199 supplemented with hormones (M1), TCM-199 supplemented with epidermal growth factor (M2) and m-SOF (M3) overlaid with paraffin oil before incubating under 5 % CO 2 in air at 38.5 o C. A part of COCs was fixed to determine the stages of nuclear maturation after 24 hr. In exp 2 the matured oocytes were allotted with sperm suspension before incubating under 5 % CO 2 in air at 38.5 o C, for 22-24 hr. Presumptive embryos were placed, individually in 96 wells petri dish to determine at 12 hr intervals the kinetics of cleavage up to blastocyst stage. Average of collected COCs per ovary was 2.4 with 86 % of excellent and good graded COC's. M1 and M2 showed higher (P<0.05) maturation rate vs. M3. After culturing oocyte diameter and perivitalline space increased (P<0.05) relative to preculture diameters, while diameter of ooplasm and thickness of zona pellucida non significantly decreased. Fertilized ova and cleaved embryos (%) were 80.2 and 77.8 %, respectively. Early embryonic cleavage up to 32 cell stage lapsed took 84 hr, while corresponding periods to reach morula, and blastocyst stage were 101 and 134.5 hr, respectively. Out of the fertilized ova 31.5 % reached excellent and good graded blastocyst stage.

Egyptian Journal of Animal Production, 2019

The current study was done to investigate the expression profile of selected candidate genes in c... more The current study was done to investigate the expression profile of selected candidate genes in corpus luteum of cyclic Egyptian buffaloes. A total number of nine corpora lutea (CLs) was collected from 9 cyclic slaughtered female Egyptian buffaloes. Based on their developmental phases and morphological features, CLs were classified into; growing (around days 3-5), static (around days 7-10) and regressed (around days 19-21 of the estrous cycle). Transcript abundance of genes regulating mitochondrial activity (SOD2, TFAM and CPT2), lipid metabolism (SREBP1), pro-inflammatory cytokine (TNFα) and oxidative stress (NFE2L2) was assessed relative to GAPDH as a housekeeping gene. Relative gene expression profiles of TFAM and SOD2 increased (P ≤ 0.05) in static phase compared to both growing and regressed phases. Transcript abundance of both CPT2 and SREBP1 was significantly higher at static phase, while was at minimum level of expression during regression phase. The expression profile of NFE2L2 revealed insignificant differences among the studied phases of CLs. Relative transcript abundance of TNFα was higher (P ≤ 0.05) at regression phase compared to growing and static ones. Taken together, the pattern of genes regulating mitochondrial activity and lipid metabolism were higher during growing phase and reached a plateau in static phase and fully declined during regression phase. Meanwhile, the transcript abundance of pro-inflammatory cytokine gene denoted an opposite trend. Thus, gene expression profile reflected the cyclic changes of buffalo corpus luteum development.

Canadian Journal of Animal Science, 2016

Feeding ruminants with Pittosporum undulatum can adversely affect the immune, endocrine and repro... more Feeding ruminants with Pittosporum undulatum can adversely affect the immune, endocrine and reproductive systems. The aim of the present study was to evaluate how feeding with P. undulatum alters progesterone levels during the oestrus cycle, and fertilization and development of oocytes after in vitro fertilization (IVF). Heifers were fed a diet containing (n=4) or not containing (n=4) this plant. Peripheral blood samples were collected every three days and progesterone levels were analysed by ELISA. The heifers' ovaries were recovered after slaughter and the oocytes were removed, maturated, in vitro fertilized and cultured for seven days, during which time their development was evaluated every two days. Results indicated that plasma concentrations of progesterone during the oestrus cycle were significantly higher (p <0.01) in the luteal and follicular phases of the animals fed without Pittosporum when compared with those fed with Pittosporum. Statistical differences (p <0.05) in maturation, cleavage rate and embryonic development were observed between oocytes belonging to the experimental and control group. Rates of maturation, fertilisation and embryo development in the experimental group were, respectively, 78.3±5.8, 29.92±4.31 and 7.30±3.1 as compared to 90.5±3.0, 41.86±58 and 21.88±6.85 in the control group. These differences can be explained as compounds from this plant can directly or indirectly inactivate cyclooxygenase 2 (COX-2), which may diminish follicular development through the inhibition of prostaglandins synthesis, oocyte maturation and, consequently, reduce the ability of oocytes to be fertilized and develop.

Zygote (Cambridge, England), Jan 8, 2017

The exposure of oocytes to heat stress during the maturation process results in harmful effects t... more The exposure of oocytes to heat stress during the maturation process results in harmful effects to their internal organelles, low fertilization capability and higher embryonic losses. In the present experiment the effect of heat shock (HS) during the maturation process was assessed. In Assay 1, oocytes from winter (December-March; n = 100) and summer (June-September; n = 100) months were collected and matured to analyse their HS tolerance. Total RNA was extracted from matured oocytes and cDNA synthesis was performed, followed by qPCR for selected genes (Cx43, CDH1, DNMT1, HSPA14), compared with two reference genes (GAPDH and SDHA). In Assay 2, oocytes collected during the winter were subjected to kinetic HS by stressing them at 39.5°C for 6, 12, 18 or 24 h and were afterwards matured at control temperature (38.5°C), and subsequently subjected to the previously described gene analysis procedure. Results of Assay 1 show that summer-collected oocytes exhibited lower maturation rate tha...