Theriogenology
Mohammadhamed Shahsavari; Gholamali Moghaddam; Kele Amaral Alves; Benner Geraldo alves; Laritza Ferreira de Lima; Golshan Azimi; Deysi jouana Dipaz Berrocal; Lucina Mascena Silva; Yago Pinto da Silva; Diego Alberto Montano Vizcarra; José Richardo de Figuereido; Ana Paula Ribeiro Rodrigues
Articles in Press, Corrected Proof, Available Online from 08 August 2023
Abstract
Although cryopreservation of ovarian tissue has advanced greatly, it remains a challenge, and protocols should be optimized to handle the heterogeneous nature of ovarian samples. In an effort to address this factor, the present study evaluated the effects of corpus luteum (CL) and side of ovaries (right ...
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Although cryopreservation of ovarian tissue has advanced greatly, it remains a challenge, and protocols should be optimized to handle the heterogeneous nature of ovarian samples. In an effort to address this factor, the present study evaluated the effects of corpus luteum (CL) and side of ovaries (right versus left) on cellular morphology and viability of vitrified bovine ovarian fragments in a closed system. The ovaries were categorized according to whether they had a CL and which side they were on, and then divided into six groups: 1) CL+ (with CL) group; 2) CL˗ (without CL) group; 3) right ovaries group; 4) left ovaries group; 5) fresh control group (ovaries without vitrification or culture that were not selected for CL or ovarian side) and 6) In vitro culture medium control group (non-vitrified ovaries that were not selected for the presence or absence of CL or side of the ovaries). The current study shows that the CL˗ and right groups had the greatest percentage of follicles with normal morphology compared to other vitrified-warmed groups. Furthermore, the levels of necrosis and tissue damage of the right cultured group were the lowest compared to other groups. It was shown that bovine ovarian tissues derived from right ovaries and ovaries without a corpus luteum can be functionally and morphologically preserved after vitrification. For the first time, the present study suggests that bovine ovarian tissue vitrification can be improved by considering the origin of the ovaries.
Theriogenology
Mohammad Bolghanabadi; Hamideh Salari Sedigh; Pezhman Mirshokraei; Masoud Rajabioun
Articles in Press, Corrected Proof, Available Online from 08 August 2023
Abstract
This study designed a protocol that would combine pregnant mare serum gonadotrophin (PMSG) and cabergoline (CAB) to induce estrus in bitches. Twenty clinically healthy adult and anestrous female dogs were randomly assigned into four groups. The first group was treated with 5.00 μg kg-1 CAB until the ...
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This study designed a protocol that would combine pregnant mare serum gonadotrophin (PMSG) and cabergoline (CAB) to induce estrus in bitches. Twenty clinically healthy adult and anestrous female dogs were randomly assigned into four groups. The first group was treated with 5.00 μg kg-1 CAB until the onset of proestrus or for 25 days. The second group was treated with 20.00 IU kg-1 PMSG for 5 days and 500 IU human chorionic gonadotropin (hCG) on the 5th day. The third group was treated with 5.00 μg kg-1 CAB for 10 days in combination with 20.00 IU kg-1 PMSG for 5 days and 500 IU hCG on the 10th day. The control group received 1.00 mL of normal saline. Ovarian changes were evaluated ultrasonographically, and the estrus cycle phase was examined by vaginal cytology. Respectively, three, three and four bitches showed clinical signs of proestrus in each treatment group. The intervals between treatment and proestrus for each group were 30.00 ± 3.05, 7.67 ± 1.20 and 13.00 ± 1.20 days, respectively. Two weeks after estrus, the progesterone mean was 14.51 ± 6.24, 19.96 ± 17.16 and 19.12 ± 9.26 ng mL-1 for each group, respectively. In ultrasonography examination, the largest follicle was identified at 15.66 ± 1.33, 11.66 ± 2.40 and 8.75 ± 2.17 days after the onset of proestrus and the largest follicle’s size was measured 6.50 ± 0.55, 4.83 ± 1.64 and 7.07 ± 1.49 mm for each group, respectively. Although the combined use of CAB and PMSG reduced the duration of treatment, alteration of the duration or PMSG dosage can be helpful to improve the results.
Theriogenology
Sadigheh Pashapour; Adel Saberivand; Amir Afshin Khaki; Maryam Saberivand
Volume 14, Issue 11 , November 2023, , Pages 601-606
Abstract
About a third of human infertility is related to male factors. Of these, idiopathic-related infertility is not curable. Diabetes mellitus is a metabolic disorder affecting male impotence and fertility by increased production of free radicals and oxidative stress. Saponin, a glycosidic compound found ...
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About a third of human infertility is related to male factors. Of these, idiopathic-related infertility is not curable. Diabetes mellitus is a metabolic disorder affecting male impotence and fertility by increased production of free radicals and oxidative stress. Saponin, a glycosidic compound found in many plants, improves sperm parameters. The present study investigated the effect of saponin on sperm oxidative stress and testicular structure in streptozotocin (STZ)-induced diabetic mice. The diabetes was induced by the administration of 150 mg kg-1 STZ via a single intra-peritoneal injection. All experimental mice were allocated to the following groups: Control group, diabetic control group, diabetic group administrated 100 mg kg-1 saponin daily and one healthy group administrated saponin daily for 56 days. At the end of the treatment period, serum levels of insulin, glucose and oxidative stress markers were measured. A histological evaluation of testicles was performed. Treatment of diabetic mice with saponin ameliorated testicular tissue damage as well as serum glucose and insulin concentrations. Furthermore, in the diabetic group, the serum concentration of malondialdehyde was increased; while, the activity of superoxide dismutase and glutathione peroxidase enzymes was reduced. The mean Johnsen's score and the diameter and thickness of seminiferous tubules were lower in the diabetic mice than control ones. However, these parameters were higher in the saponin-treated mice than controls. Overall, saponin administration rectified all examined parameters. The anti-oxidant role of saponin improves sperm parameters and diabetes-induced testicular oxidative damage.
Theriogenology
Sharareh Ahmadi; Adel Saberivand; Cyrus Jalili; Reza Asadpour; Monire Khordadmehr; Maryam Saberivand
Volume 14, Issue 9 , September 2023, , Pages 507-513
Abstract
Triple-negative breast cancer (TNBC) is an aggressive and deadly breast cancer sub-type with limited therapeutic options. Dandelion (Taraxacum officinale) exhibiting extensive anti-cancer activity is reported to be effective against TNBC; however, its anti-tumor effect mechanisms have not been fully ...
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Triple-negative breast cancer (TNBC) is an aggressive and deadly breast cancer sub-type with limited therapeutic options. Dandelion (Taraxacum officinale) exhibiting extensive anti-cancer activity is reported to be effective against TNBC; however, its anti-tumor effect mechanisms have not been fully elucidated. The purpose of this study was to determine the anti-cancer activity of hydroalcoholic extract of dandelion (HADE) on 4T1 cells, and the mechanism of HADE-induced cell death. The effect of HADE on cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and lactate dehydrogenase assays. Apoptotic cell death was monitored by flow cytometry. The DNA fragmentation was evaluated by Acridine orange/Ethidium bromide (AO/EB) staining. Nitric oxide (NO) level was detected using Griess assay. The effects of HADE on Atg-7, Beclin-1, Bcl2, Bax and p53 genes were investigated by real-time reverse transcription-polymerase chain reaction. The results showed that HADE inhibited cell growth and proliferation in a dose- and time-dependent manner. The HADE induced 4T1 breast cancer cell death via apoptosis and autophagy. The DNA fragmentation was improved as the concentration of HADE increased. The NO secretion was declined with increasing concentration of HADE. Gene expression analysis confirmed HADE-induced apoptosis and autophagy in cancer cells. The Bax, Bax/Bcl-2 ratio, p53, Beclin-1 and Atg-7 over-expression as well as Bcl-2 down-regulation were also evident in treated cancer cells.
Theriogenology
Arman Shokri; Reza Asadpour; Razi Jafai-joozani; Esmaeil Babaei; Abolfazl Hajibemani; Gholamreza Hamidian
Volume 14, Issue 8 , August 2023, , Pages 437-445
Abstract
The purpose was to identify differentially expressed plasma microRNAs (miRNAs) in cows with clinical and subclinical endometritis. In this study clinical endometritis (CE; n = 23) based on vaginal discharge score (VDS), subclinical endometritis (SCE; n = 17) based on VDS (0), and endometrial cytology ...
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The purpose was to identify differentially expressed plasma microRNAs (miRNAs) in cows with clinical and subclinical endometritis. In this study clinical endometritis (CE; n = 23) based on vaginal discharge score (VDS), subclinical endometritis (SCE; n = 17) based on VDS (0), and endometrial cytology (the presence of 8.00% polymorphonuclear neutrophils (PMN) on days 21-31 and 5.00% on days 41-51 days in milk (DIM) and healthy cows (n = 21) based on vaginal discharge score (0), and endometrial cytology (< 5.00% PMN on days 21 - 31 and < 5.00% on days 41 - 51 DIM) were selected. The results showed that the expression level of miR-146a was significantly higher in the CE (19.17-fold), and SCE (6.22-fold) groups than those of healthy cows. The relative transcript abundance of miR-223 was considerably down-regulated in the CE (0.26-fold) and SCE (0.06-fold) compared to the healthy cows. The expression levels of miR-146a and miR-223 were significantly higher in the CE group which could be caused by Gram-negative bacterial infection. Our results showed that the expression level of plasma miRNAs postpartum could be used as a reliable marker to distinguish between SCE, CE and healthy cows.
Theriogenology
Kimia Maleki; Esmail Ayen; Amir Khaki; Ali Soleimanzadeh
Volume 14, Issue 7 , July 2023, , Pages 373-379
Abstract
Sperm survives for a very short time in fresh semen, and slow cooling to 5.00 ˚C kills a large number of sperms. This study was aimed to compare the semen quality parameters and anti-oxidant levels in four extenders (manual, Triladyl, Steridyl and AndroMed). Semen samples were obtained from a total ...
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Sperm survives for a very short time in fresh semen, and slow cooling to 5.00 ˚C kills a large number of sperms. This study was aimed to compare the semen quality parameters and anti-oxidant levels in four extenders (manual, Triladyl, Steridyl and AndroMed). Semen samples were obtained from a total number of 12 dual-purpose Simmental bulls kept in the Simmental Cattle Breeding Center for a period of 3 months using an artificial vagina. Sperm viability, motility, abnormal morphology, plasma membrane integrity, DNA damage, chromatin quality, total antioxidant capacity (TAC) and lipid peroxidation were evaluated. The highest progressive motility, viability, plasma membrane integrity, and TAC and the lowest levels of malondi-aldehyde in the frozen-thawed semen belonged to the semen group frozen with Triladyl. Parameters of motility were higher in the frozen-thawed semen with Triladyl than in other groups, indicating a significant difference from the manual extender. Among the extenders studied, Triladyl was the most suitable for semen freezing in Simmental bulls.
Theriogenology
Héctor Javier Narváez; Luis Ernesto Quintero Rodríguez
Volume 14, Issue 7 , July 2023, , Pages 397-400
Abstract
The creole breed Blanco Orejinegro (Bos taurus adapted) is an important animal genetic resource in Colombia. However, very little is known about the reproductive physiology of the breed. The objective was to determine the dynamics of progesterone circulation and follicular growth in Blanco Orejinegro ...
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The creole breed Blanco Orejinegro (Bos taurus adapted) is an important animal genetic resource in Colombia. However, very little is known about the reproductive physiology of the breed. The objective was to determine the dynamics of progesterone circulation and follicular growth in Blanco Orejinegro cows throughout the estrous cycle. Fifteen cyclic Blanco Orejinegro cows were used and subjected to hormonal protocol for estrous and ovulation synchronization. Once the time of ovulation was identified by monitoring the ovarian dynamics with ultrasonography equipment. For description of the results, the mean was used as a measure of central tendency and the standard error of the mean. Statistical analysis was longitudinal descriptive. Blood samples were collected every 24 hr throughout the estrous cycle for serum quantification of progesterone (P4), monitoring of corpus luteum (CL) growth dynamics and follicular dynamics. It was possible to verify that 76.92% of the Blanco Orejinegro cows presented two follicular waves during the estrous cycle and the highest circulating levels of P4 (> 6.00 ng mL-1) were observed on the 14th day of the estrous cycle. It was concluded that the creole cows of the Blanco Orejinegro breed presented two follicular waves per estrous cycle. In addition to, the size of the CL was consistent with the secretion of progesterone presenting higher circulating levels at the end of the luteal phase.
Theriogenology
Amir Khaki; Atefeh Araghia; Mehdi Lotfi; Alireza Nourian
Volume 10, Issue 4 , December 2019, , Pages 333-341
Abstract
Artificial insemination is a well-established and widely used method for genetic improvement in cattle breeding industry. Recently, researchers have shown an increased interest in the cryoprotective effects of minerals and antioxidants on semen. Previous studies on calcium (Ca) and magnesium (Mg), two ...
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Artificial insemination is a well-established and widely used method for genetic improvement in cattle breeding industry. Recently, researchers have shown an increased interest in the cryoprotective effects of minerals and antioxidants on semen. Previous studies on calcium (Ca) and magnesium (Mg), two main macro-minerals, have mainly investigated their roles in mammalian spermatogenesis and fertility. In addition, the experimental data examining the semen content regarding these minerals and antioxidants from different animal species are rather controversial and there is no general agreement about their associations with semen quality. Therefore, this study was conducted to assess the seminal plasma concentrations of Ca, Mg and total antioxidant capacity (TAC) in first and second ejaculations of dual-purpose Fleckvieh bulls and to link them to the sperm characteristics of fresh and frozen-thawed semen. Sperm progressive motility after thawing was used to classify the data into three groups: < 40.00%, 40.00 to 50.00% and > 50.00%. The measurements of two minerals and TAC were carried out using spectrophotometry and enzyme-linked immunosorbent assays, respectively. The results showed that there were significant differences in several parameters of semen quality between first and second ejaculations. No significant differences were also found on Ca and Mg concentrations and Ca/Mg ratio. The TAC level was significantly higher in the first ejaculation than the second one. The findings of this study suggest that TAC is a potential marker for bull semen quality assessment in the frozen semen production industry.
Theriogenology
Parisa Aparnak; Adel Saberivand
Volume 10, Issue 3 , September 2019, , Pages 221-226
Abstract
Canine seminal plasma contains antioxidant enzymes to protect sperm against internally generated ROS. These enzymes are removed from seminal plasma during the process of cryopreservation. The freezing/thawing process can cause some morphological and functional changes via ice crystallization and osmolality ...
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Canine seminal plasma contains antioxidant enzymes to protect sperm against internally generated ROS. These enzymes are removed from seminal plasma during the process of cryopreservation. The freezing/thawing process can cause some morphological and functional changes via ice crystallization and osmolality imbalance. The present study was conducted to evaluate the effects of curcumin supplementation on sperm total count, motility, progressive motility, viability, morphology, total antioxidant capacity (TAC), DNA integrityand NOX5 gene expression of dog frozen semen. The pooled semen was allocated to fresh (Group 1) and frozen (Group 2) controls, curcumin (2.50 mM) (Group 3) and curcumin (5.00 mM), (Group 4). Sperm parameters including total sperm count, morphology, motility, progressive motility, sperm concentration and DNA integrity in addition to TAC were evaluated in fresh and frozen-thawed semen samples. Real-time PCR was used to investigate NOX5 and GADPH (reference gene) genes expressions. Curcumin at 2.50 mM provided a greater protective effect on the DNA integrity compared to 5.00 mM and control groups. TAC was significantly higher in 2.50 mM group than other groups. NOX5 gene expression in curcumin 2.50 mM was higher than 5.00 mM group. In conclusion, curcumin seems to emolliate sperm parameters and to protect sperm against sperm reactive oxygen stress and increases NOX5 gene expression.
Theriogenology
Siamak Kazemi-Darabadi; Reza Asadpour; Amir Ali Shahbazfar; Sajjad Alizadeh
Volume 10, Issue 2 , June 2019, , Pages 125-132
Abstract
Testicular torsion is a consequence of spermatic cord twisting which causes progressive damage to the structure of the testis and reduces sperm quality and usually results in infertility. In the present study, with the assumption of the protective effects of L-carnitine and betamethasone against ischemia-reperfusion ...
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Testicular torsion is a consequence of spermatic cord twisting which causes progressive damage to the structure of the testis and reduces sperm quality and usually results in infertility. In the present study, with the assumption of the protective effects of L-carnitine and betamethasone against ischemia-reperfusion (IR) injuries, their effects on twisted testicles were evaluated and compared. Twenty Wistar rats were randomly divided into four groups and used in this study. Except for the Sham (S) group, testicular IR was induced surgically in three other groups, including Control (C), Betamethasone (BM), and L-carnitine (LC) groups. Betamethasone and L-carnitine were injected before detorsion in the BM and LC groups, respectively. After twelve hours of reperfusion, the testicles were detached, and prepared for sperm parameters evaluation such as sperm count, motility, viability, morphology, and chromatin quality, and histopathologic evaluations, including mean seminiferous tubular diameter (MSTD), germinal epithelial cell thickness (GECT), and Johnsen’s mean testicular biopsy scoring (MTBS). The MSTD, GECT, and healthy sperms in the C group were significantly lower than the other groups, while the BM and LC groups were significantly different from others in MTBS. The number of sperms and sperm motility in the BM group was significantly higher than the C group. Sperm viability in the BM and LC groups were significantly higher than the C group. The results of this study showed that both L-carnitine and betamethasone similarly can be effective in treating testicular IR injuries.
Theriogenology
Abolfazl Mohammadi; Hesam Seifi; Nima Farzaneh
Volume 10, Issue 2 , June 2019, , Pages 153-158
Abstract
The present study aimed to determine the effect of administrating prostaglandin F2α (PGF2α) and GnRH at the time of artificial insemination (AI) on the pregnancy per artificial insemination (P/AI) and the pregnancy survival rate of dairy cows. A number of 830 lactating Holstein cows were ...
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The present study aimed to determine the effect of administrating prostaglandin F2α (PGF2α) and GnRH at the time of artificial insemination (AI) on the pregnancy per artificial insemination (P/AI) and the pregnancy survival rate of dairy cows. A number of 830 lactating Holstein cows were randomly divided into four groups. Cows in group 1 (n=200) treated with 150 µg d-cloprostenol. In group 2 (n=212), cows received 10 µg buserelin acetate, and group 3 (n=205) was treated with both 150 µg d-cloprostenol and 10 µg buserelin acetate. In addition, 213 cows were assigned as control group which received normal saline as placebo (group 4). To measure progesterone, milk samples were collected at the insemination day and five days later. Pregnancy diagnosis was performed 28 and 60 days after the insemination, and the size and number of corpus luteum (CL) and twin pregnancies were recorded. Hormone therapies had no effect on the P/AI, pregnancy survival rate, and the size and number of CL. The P/AI ratio in groups 1, 2, 3 and 4 were 38.50%, 42.92%, 41.46% and 40.84%, and the pregnancy survival rates in groups 1, 2, 3 and 4 were 84.42%, 86.81%, 88.23% and 83.91%, respectively. The probability of a twin pregnancy was significantly higher in group 1 (15.58%) than other groups. There was no significant difference between groups in terms of the offspring gender. In conclusion, the administration of d-cloprostenol or buserelin acetate at the time of AI had no effect on P/AI and pregnancy survival rate in dairy cattle under no heat stress condition, while the administration of d-cloprostenol increased the probability of twin pregnancies.
Theriogenology
Reza Narenji Sani; Bahram Hajigolikhani; Mahmood Ahmadi-Hamedani; Khatereh Kafshdouzan
Volume 9, Issue 4 , December 2018, , Pages 343-348
Abstract
The aim of the present study was to investigate the diagnostic evaluation of milk lactate dehydrogenase (mLDH) and alkaline phosphatase (mALP) activities by receiver operating characteristic (ROC) analysis curve in early lactation of ewes with subclinical mastitis (SCM) and determine the correlation ...
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The aim of the present study was to investigate the diagnostic evaluation of milk lactate dehydrogenase (mLDH) and alkaline phosphatase (mALP) activities by receiver operating characteristic (ROC) analysis curve in early lactation of ewes with subclinical mastitis (SCM) and determine the correlation between number of somatic cell count (SCC) and mLDH and mALP activities. A total of 196 udder half milk samples were collected within the first 6 weeks of lambing. The SCM was determined by positive milk bacterial culture and positive California mastitis test (CMT); SCC was determined by fossomatic method and enzyme activities were determined spectrophotometrically. The mLDH and mALP of SCM cases were positively correlated with SCC values. Values of mLDH, mALP and SCC were significantly higher in SCM than non-SCM udder halves. The optimum cut-off points of mLDH and mALP activities for SCM diagnosis were determined at 203.61 (U L-1) and 329.84 (U L-1), respectively. In conclusion, SCC has positive correlation with mALP and mLDH activities in SCM ewes and mLDH and mALP activities could be considered as reliable indicators for intra-mammary inflammation diagnosis.
Clinical Pathology
Alireza Kabirian; Rooz Ali Batavani; Siamak Asri-Rezaei; Ali Soleimanzadeh
Volume 9, Issue 3 , September 2018, , Pages 217-224
Abstract
Cyclophosphamide is a chemotherapy drug for the treatment of cancer. Chicken embryo amniotic fluid, vitamin C and coenzyme Q10 have anti-oxidant properties. Total of 70 adult female mice were selected and divided into seven groups. The first group that received 2 ml kg-1 of inactivated amniotic fluid ...
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Cyclophosphamide is a chemotherapy drug for the treatment of cancer. Chicken embryo amniotic fluid, vitamin C and coenzyme Q10 have anti-oxidant properties. Total of 70 adult female mice were selected and divided into seven groups. The first group that received 2 ml kg-1 of inactivated amniotic fluid subcutaneously. The second group treated with 75 mg kg-1of cyclophosphamide by intraperitoneal injection. Third to fifth groups received 1, 2, and 4 ml kg-1 of chicken embryo amniotic fluid, respectively. The sixth group received vitamin C at a dose of 0.2 mg g-1 of body weight by oral gavages. Seventh group received 10 mg kg-1 coenzyme Q10 intraperitoneally. All cyclophosphamide treated groups (3-7) received 75 mg kg-1 of cyclophosphamide intraperitoneal on day 22. The mice were euthanized on day 29 and ovarian tissue antioxidant enzymes including glutathione peroxidase, superoxide dismutase and catalase activities and malondialdehyde (MDA) were evaluated. Activities of above mentioned enzymes in treatment groups (3-7) was significantly higher than patient control group (2). The results also revealed that MDA levels were higher in the control group in comparison to other treatment groups. Therefore, it is concluded that the chick embryo amniotic fluid and coenzyme Q10 can compete with compounds like vitamin C in increasing the anti-oxidant level in ovarian tissue.
Theriogenology
Ali Soleimanzadeh; Leila Mohammadnejad; Abbas Ahmadi
Volume 9, Issue 3 , September 2018, , Pages 265-271
Abstract
Busulfan is known to cause several adverse effects including reproductive toxicity in humans. Garlic (Allium sativum), a widely distributed medicinal plant, is highly regarded for its medicinal activities including antioxidant property.This study was conducted to assess whether garlic extract could serve ...
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Busulfan is known to cause several adverse effects including reproductive toxicity in humans. Garlic (Allium sativum), a widely distributed medicinal plant, is highly regarded for its medicinal activities including antioxidant property.This study was conducted to assess whether garlic extract could serve as protective agents against testicular toxicity during busulfan treatment in a mice model.Seventy-two adult male mice were randomly divided into nine groups. In groups 1,2 and 3, distilled water, busulfan, and dimethyl sulfoxide and in the treatment groups hydro-alcoholic extract of garlic was administered orally at different doses per day (groups 4, 5 and 6; 200, 400, 800 mg kg-1 respectively). Groups 7, 8 and 9 were treated with the extract (200, 400 and 800 mg kg-1, respectively) plus busulfan. Following euthanasia, blood samples and epididymal sperm were collected.The busulfan-treated group showed significant decreases in sperm qualityparameters, and serum levels of testosterone, LH and FSH was observed in the busulfan-treated mice. In addition, the TAC levels and antioxidant enzymes activities were reduced and malondialdehyde (MDA) levels were increased in the busulfan-treated mice. Notably, garlic extract co-administration caused a considerable recovery in sperm qualityparameters, TAC levels, antioxidant enzymes activities, hormonal changes and MDA level. Based on our results, garlic has antioxidant effects against busulfan-induced testicular damages in mice.
Theriogenology
Mahboobeh Amoushahi; Mojdeh Salehnia
Volume 9, Issue 2 , June 2018, , Pages 145-152
Abstract
The aim of this study was to evaluate the effects of ovarian tissue vitrification and two-step in vitro culture on the metaphase II (MII) oocyte reactive oxygen species (ROS) level, mitochondrial transcription factor A (TFAM) expression and succinate dehydrogenase (SDH) activity. After collection of ...
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The aim of this study was to evaluate the effects of ovarian tissue vitrification and two-step in vitro culture on the metaphase II (MII) oocyte reactive oxygen species (ROS) level, mitochondrial transcription factor A (TFAM) expression and succinate dehydrogenase (SDH) activity. After collection of neonatal mouse ovaries, 45 ovaries were vitrified and the others (n = 45) were considered as control. All ovaries were cultured for seven days, and their isolated preantral follicles were cultured in three-dimensional culture system. After 12 days in vitro culture, the follicular development and oocyte maturation were evaluated and compared in vitrified and non-vitrified ovaries. The collected MII oocytes were inseminated with capacitated spermatozoa. The fertilization, embryonic development, ROS level, TFAM gene expression and SDH activity of oocytes were assessed and compared. There was no significant difference between morphology and percentage of normal follicles between vitrified and non-vitrified ovaries at the beginning of culture. The follicular development and hormone level in the vitrified group was significantly lower than non-vitrified group and the ROS concentration in the vitrified group was significantly higher than non-vitrified group after one-week organ culture. After follicular culture, there was no significant difference in follicular development, oocyte maturation, fertilization rate, TFAM gene expression, ROS level and mitochondrial SDH activity between vitrified and non-vitrified groups. This study showed that mouse ovarian tissue vitrification influenced the follicular development through increase in ROS level during organ culture but these harmful effects of vitrification method may be recovered during the follicular culture period. Thus, vitrification and ovarian organ culture method should be improved.
Theriogenology
Soghra Abbaszadeh; Farhad Farrokhi-Ardabili; Hassan Malekinejad; Iraj Bernousi
Volume 9, Issue 2 , June 2018, , Pages 171-178
Abstract
This study was aimed to investigate the effects of 17 𝛽-estradiol (E2) and 𝛼-zearalenol (α-ZOL) on motility parameters, plasma membrane integrity, levels of produced nitric oxide (NO) and total antioxidant capacity of Ghezel ram sperm during the liquid storage at 4 ˚C, for various periods ...
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This study was aimed to investigate the effects of 17 𝛽-estradiol (E2) and 𝛼-zearalenol (α-ZOL) on motility parameters, plasma membrane integrity, levels of produced nitric oxide (NO) and total antioxidant capacity of Ghezel ram sperm during the liquid storage at 4 ˚C, for various periods of time. Semen samples were collected from four rams and diluted with Tris–egg yolk extender and supplemented with E2 (100 µmol) or different concentrations of α-ZOL (100 pmol, 100 nmol and 100 µmol) at a final concentration of 200 × 106 sperm per mL. We failed to show any significant effect of E2 at 100 µmol concentration on ram’s sperm parameters while α-ZOL resulted in a significant decrease of plasma membrane integrity at 100 µmol concentration (55.40% for α-ZOL vs 62.20% for control) after 96 hr incubation. Alpha-ZOL had decreasing effect on sperm motility parameters including curvilinear velocity and average path velocity at 100 µmol concentration after 96 hr storage. Although remarkable reduction of total antioxidant capacity at high concentration of α-ZOL and long incubation time was found, however no significant changes were recorded in NO level during storage time. It was concluded that the detrimental effect of α-ZOL on ram sperm might be attributed to its induced oxidative stress and damage to the plasma membrane.
Theriogenology
Somayeh Tavana; Mojtaba Rezazadeh Valojerdi; Hussein Eimani; Naeimeh Sadat Abtahi; Rouhollah Fathi
Volume 8, Issue 4 , December 2017, , Pages 275-280
Abstract
This study was carried out to assess the different ovarian transplantation sites after short-time autografting. Female rats were randomized into five groups, with six rats in each group, including control (intact), cervical subcutaneous transplanted (CST), back subcutaneous transplanted (BST), subfascial ...
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This study was carried out to assess the different ovarian transplantation sites after short-time autografting. Female rats were randomized into five groups, with six rats in each group, including control (intact), cervical subcutaneous transplanted (CST), back subcutaneous transplanted (BST), subfascial transplanted (SFT) and intramuscular transplanted (IMT) groups. In all experimental groups, the right ovary was removed and transplanted into different sites. After three weeks, ovaries were removed for morphology assessment, follicular counting and the rates of corpus luteum (CL) and cyst formation. Transplanted ovaries in BST and SFT groups were full of cysts and did not have sufficient numbers of intact follicles and were excluded from experiments. In IMT and CST groups, re-anastomosis, follicular development and good homogeneity of the stromal tissue were seen. However, the difference in intact antral follicles between CST (7.92 ± 0.02%) and CST-Op (opposite ovary of CST group) (30.99 ± 0.03%) was significant as well as the difference between CST (7.92 ± 0.02%) and control (10.08 ± 0.01%) groups. In addition, the number of intact primordial follicles in the CST-Op (16.58 ± 0.02%) group was significantly less than that of the control (40.40 ± 0.03%) group. Interestingly, the number of CL was significantly increased in the CST-Op (11.71 ± 0.01%) and IMT-Op (9.16 ± 0.02%) groups compared to the control and experimental groups. Although both intramuscular and subcutaneous sites effectively preserved ovarian follicles after three weeks, cervical subcutaneous site was better suited for auto-transplantation in rat.
Pharmacology
Abbas Ahmadi; Saleh Bamohabat Chafjiri; Rajab Ali Sadrkhanlou
Volume 8, Issue 4 , December 2017, , Pages 281-286
Abstract
Busulfan is an alkylating agent affects ovarian follicles growth by oxidative stress induction. Satureja khuzestanica has antioxidant effects. The aim of this study was to examine whether S. khuzestanica essential oil (SKEO) exhibits protective effects on busulfan-induced ovarian failure. Eighty-four ...
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Busulfan is an alkylating agent affects ovarian follicles growth by oxidative stress induction. Satureja khuzestanica has antioxidant effects. The aim of this study was to examine whether S. khuzestanica essential oil (SKEO) exhibits protective effects on busulfan-induced ovarian failure. Eighty-four adult female mice were divided into six groups including dimethyl sulfoxide (control), SKEO 225.00 mg kg-1 (orally), busulfan 3.00 mg kg-1 (orally), busulfan 36.00 mg kg-1 (intraperitoneally), busulfan 3.00 mg kg-1 and SKEO and busulfan 36.00 mg kg-1 and SKEO. After 28 days, the mice were euthanized and oocytes were removed for in vitro fertilization (IVF) rate evaluation. Oocyte quantity and quality, fertilization rate and pre-implantation embryo development were daily examined with a stereo microscope in a period of 120 hr. Serum levels of estradiol and progesterone were also evaluated. Busulfan caused significant decreases in oocyte number and quality, fertilization rate, pre-implantation embryo development and embryo quality. The SKEO significantly decreased the adverse effects of busulfan. The present study indicated that SKEO can protect female fertility potential against busulfan induced damages.
Pharmacology
Alireza Nourian; Ali Soleimanzadeh; Ali Shalizar Jalali; Gholamreza Najafi
Volume 8, Issue 4 , December 2017, , Pages 341-345
Abstract
Bisphenol-S (BPS) is a new bisphenol-A substitute widely used in many plastic products. Bisphenol-A as a main member of bisphenol family has been known as an endocrine system disrupter chemical compound. Like other members of bisphenol family, there is public health concern about the toxic effects of ...
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Bisphenol-S (BPS) is a new bisphenol-A substitute widely used in many plastic products. Bisphenol-A as a main member of bisphenol family has been known as an endocrine system disrupter chemical compound. Like other members of bisphenol family, there is public health concern about the toxic effects of BPS on reproductive system, thus, we examined BPS effects on in vitro fertilization (IVF) potential and oxidative stress status in a murine model. Adult female mice (n = 70) were randomly divided into control and BPS-treated groups. Bisphenol-S was administered at doses of 0, 1, 5, 10, 50 and 100 µg kg-1 body weight per day intraperitoneally for 21 consecutive days. Twenty-Four hr after the last treatment, five mice in each group were super-ovulated and the oocytes were harvested for IVF. All ovaries were collected and used for biochemical factors analyses. Bisphenol-S exposure at doses more than 10 µg kg-1 induced developmental arrest of pre-implantation embryos. Further, lipid peroxidation measurement in ovaries indicated that all doses of BPS cause oxidative stress in female mice. In conclusion, BPS administration even in low doses can result in female reproductive toxicities and oxidative stress in mice.
Theriogenology
Hossein Hassanpour; Valiallah Khalaji-Pirbalouty; Manoochehr Adibi; Hassan Nazari
Volume 8, Issue 3 , September 2017, , Pages 251-257
Abstract
Peroxisome proliferator-activated receptors (PPARs) are nuclear receptors of transcription factors composed of three family members: PPARα, PPARβ/δ and PPARγ. This study was aimed to evaluate the role of PPARs in the estradiol production via follicle stimulating hormone (FSH) in ...
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Peroxisome proliferator-activated receptors (PPARs) are nuclear receptors of transcription factors composed of three family members: PPARα, PPARβ/δ and PPARγ. This study was aimed to evaluate the role of PPARs in the estradiol production via follicle stimulating hormone (FSH) in the ovine Sertoli cells. At the first step, transcripts of PPARα, PPARβ /δ and PPARγ were evaluated by quantitative real time PCR (qRT-PCR) in the ovine Sertoli cells in vitro after FSH treatment. PPARγ transcript was increased in FSH-treated cells while PPARα and PPAR β /δ transcripts were unchanged. At the second step, Pioglitazone as PPARγ agonist and 2-chloro-5-nitrobenzanilide (GW9662) as PPARγ antagonist were used in the FSH-treated Sertoli cells and then, the estradiol production and aromatase transcript were evaluated. Aromatase transcript was increased by pioglitazone in the FSH-treated Sertoli cells while GW9662 did not change its transcript. The estradiol production was increased by low concentrations of pioglitazone in FSH-treated Sertoli cells while the production of this hormone was decreased by the high concentration of Pioglitazone. The GW9662 did not change the production of estradiol in FSH-treated Sertoli cells. It is concluded that FSH regulates the estradiol production and aromatase expression in a way independently of PPARβ/δ and PPARα activation, although FSH increases the transcript of PPARγ and in this way, it could affect (mostly increase) aromatase transcript and estradiol production. Probably, this effect of FSH in the estradiol production via PPARγ is only a servo-assist mechanism which if it was inhibited, the estradiol production was not considerably affected.
Parasitology
Maryam Ansari-Lari; Abbas Rowshan-Ghasrodashti; Hadi Jesmani; Maliheh Masoudian; Mehrdad Badkoobeh
Volume 8, Issue 2 , June 2017, , Pages 109-114
Abstract
There is no prospective study from Iran to estimate the direct risk of Neospora caninum for pregnancy loss or reproductive factors. In addition, there is no report in the literature concerning the association of N. caninum with dystocia and sex of calves. Therefore, this study was conducted on a group ...
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There is no prospective study from Iran to estimate the direct risk of Neospora caninum for pregnancy loss or reproductive factors. In addition, there is no report in the literature concerning the association of N. caninum with dystocia and sex of calves. Therefore, this study was conducted on a group of dairy cows in a large intensive production system during 2011 to 2013 in southern Iran to evaluate the impact of neosporosis on reproductive performance. A total of 253 cows which were diagnosed as pregnant during the first six months of the study were followed until calving or abortion. Reproductive data were collected and N. caninum serostatus was determined using ELISA. To investigate the association of abortion with N. caninum,survival analysis was performed using Cox proportional hazard model. The N. caninum seroprevalence in the study group was 30.40% (95% CI: 27.40, 36.10). The overall abortion rate was 12.25%, significantly higher in seropositive animals (20.80%) than seronegative ones (8.50%). Results of Cox model showed that serostatus of animal for N. caninum and season had significant associations with abortion (p < 0.01). Neospora caninum did not show significant association with other factors such as dystocia and sex of calves. In conclusion, neosporosis is responsible for 12.00% excess abortion risk in infected group and more than 30.00% of abortions could be preventable by control of Neospora in study population. Therefore, control of N. caninum would reduce the economic losses caused by parasite mainly due to pregnancy loss.
Theriogenology
Clifford Nwabugwu Abiaezute; Innocent Chima Nwaogu; Udensi Maduabuchi Igwebuike
Volume 8, Issue 1 , March 2017, , Pages 1-6
Abstract
The objective of this study was to highlight the postnatal development of the uterine tubes of the West African Dwarf goat from birth to 28 weeks of age by gross examination and light microscopy. There was a caudal migration of the paired uterine tubes from behind the paired kidneys at birth to the pelvic ...
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The objective of this study was to highlight the postnatal development of the uterine tubes of the West African Dwarf goat from birth to 28 weeks of age by gross examination and light microscopy. There was a caudal migration of the paired uterine tubes from behind the paired kidneys at birth to the pelvic inlet at week 8 of age. Each uterine tube exhibited three segments namely; infundibulum, ampulla and isthmus. A marked flexure, the utero-tubal junction, was the point at which the uterine tubes joined the uterine horns. The length and absolute weight of the uterine tubes increased from 4.95 ± 0.28 cm and 0.02 ± 0.01 g at birth to 14.98 ± 2.79 cm and 0.22 ± 0.03 g at week 28 of age, respectively. The mucosa of the infundibulum and the ampulla showed long, branched and anastomosing primary, secondary and tertiary mucosal folds which decreased in height towards the isthmus. The mucosal folds within the isthmus were short and lacked the anastomosing pattern. The epithelia of all three segments were pseudostratified columnar. Numerous secretory blebs and extruded nuclei became apparent from week 16 of age. The thickness of the tunica muscularis varied with the segments.
Stem Cells
Vahid Akbarinejad; Parviz Tajik; Mansoureh Movahedin; Reza Youssefi
Volume 8, Issue 1 , March 2017, , Pages 7-13
Abstract
Testosterone is believed to play a significant role in spermatogenesis, but its contribution to the process of spermatogenesis is not completely understood. Given that extracellular matrix (ECM) facilitates differentiation of spermatogonial stem cells (SSCs) during culture, the present study was conducted ...
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Testosterone is believed to play a significant role in spermatogenesis, but its contribution to the process of spermatogenesis is not completely understood. Given that extracellular matrix (ECM) facilitates differentiation of spermatogonial stem cells (SSCs) during culture, the present study was conducted to elucidate whether testosterone contribute to the permissive effect of ECM on SSCs differentiation. In experiment 1, testosterone production was measured in testicular cells cultured for 12 days on ECM or plastic (control). In experiment 2, testosterone production was assessed in testicular cells cultured on ECM or plastic (control) and exposed to different concentrations of hCG. In experiment 3, the gene expression of factors involved in testosterone production was analyzed. Testosterone concentration was lower in ECM than in the control group in experiment 1 (p < 0.05). In experiment 2, testosterone concentration was increased in response to hCG in both groups but cells cultured on ECM were more responsive to hCG than those cultured on plastic (p < 0.05). In the experiment 3, qRT-PCR revealed the inhibitory effect of ECM on the gene expression of steroidogenic acute regulatory protein (StAR) (p < 0.05). Nevertheless, the expression of LH receptor was greater in ECM-exposed than in unexposed cells (p < 0.05). In conclusion, the present study showed that inhibiting the expression of StAR, ECM could lower testosterone production by Leydig cells during in vitro culture. In addition, the results indicated that ECM could augment the responsiveness of Leydig cells to hCG through stimulating the expression of LH receptor.
Theriogenology
Narges Bagheripour; Saeed Zavareh; Mohammad Taghi Ghorbanian; Seyed Hassan Paylakhi; Seyed Reza Mohebbi
Volume 8, Issue 1 , March 2017, , Pages 43-48
Abstract
The transcriptional factor OCT4 regulates pluripotency of stem cells and has an important role during oocyte growth. Whereas, its role has remained ambiguous in ovarian tissue during reproductive cycle. Therefore, this study was aimed to investigate the expression patterns of OCT4 in mouse ovaries during ...
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The transcriptional factor OCT4 regulates pluripotency of stem cells and has an important role during oocyte growth. Whereas, its role has remained ambiguous in ovarian tissue during reproductive cycle. Therefore, this study was aimed to investigate the expression patterns of OCT4 in mouse ovaries during the normal estrous cycle. Adult National Medical Research Institute mice were classified as proestrous, estrous, metestrous and diestrous on the basis of vaginal smear cytology. Their ovaries were removed and the protein and gene expression levels of OCT4 were assessed using immunohistochemical staining and real-time quantitative reverse-transcription PCR, respectively. Immunohistochemical staining revealed the expression of OCT4 in the cytoplasm of corpus luteum cells. In the follicles, OCT4 was expressed in the cytoplasm of granulosa cells. Furthermore, the gene expression levels of OCT4 was significantly higher in the proestrous phase than in the other phases of the estrous cycle (p < 0.05). The results indicated that OCT4 gene expression levels are affected by the cyclic pattern of the estrous cycle.
Theriogenology
Reza Youssefi; Parviz Tajik; Mansoureh Movahedin; Vahid Akbarinejad
Volume 7, Issue 4 , December 2016, , Pages 275-280
Abstract
Enrichment of cell suspension with germ cells prior to injection into recipient seminiferous tubules is of importance in spermatogonial stem cells (SSCs) transplantation. Knock-out serum replacement (KSR) has been reported to enhance the proliferation of murine SSCs and human embryonic stem cells. The ...
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Enrichment of cell suspension with germ cells prior to injection into recipient seminiferous tubules is of importance in spermatogonial stem cells (SSCs) transplantation. Knock-out serum replacement (KSR) has been reported to enhance the proliferation of murine SSCs and human embryonic stem cells. The aim of the present study was to investigate the effect of KSR versus fetal bovine serum (FBS) and their interaction on colonization of bovine SSCs in vitro. When FBS (10%) was replaced with KSR (10%), a significant increase in the colonization of SSCs and the expression of Thy1, as marker for enrichment of SSCs, was observed. It was revealed that the lesser proliferative effect of FBS as well as the greater proliferative impact of KSR on SSCs colonization were not irreversible as cells having been cultured with FBS (10%) for three days with low colonization showed high rate of colonization in response to KSR (10%) and cells having been cultured with KSR (10%) with high colonization experienced low rate of colonization in response to FBS (10%). Further, it was shown that FBS did not contain factors inhibiting SSCs colonization and it simply lacked factors essential for SSCs proliferation because the combination of FBS (5%) and KSR (5%) resulted in even greater rate of colonization than did KSR (10%). In conclusion, the present study showed that addition of KSR to culture medium would significantly increase SSCs proliferation.
